count source_label source_id relationship target_label target_id entity_type solr_id publication_id sentences 26 RAD23A UNIPROT:P54725 inhibits UNIPROT:O14757 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Recovery of hHR23A expression in sh-HR23A cells reversed the stability of Chk1 to the level seen in controls (Fig. 3D).|||Indeed, knockdown of hHR23A was found to increase the protein half-life (and thus the protein level) of Chk1 (Fig. 3B and C).|||Depletion of hHR23A upregulated Chk1 at the protein level but did not alter its mRNA expression (Supplemental Fig. 1C and D).|||Depletion of hHR23A upregulated Chk1 at the protein level but did not alter its mRNA expression (Supplemental Fig. 1C and D).|||(B) The induction of Chk1 protein levels by hHR23A knockdown was not cell-context-dependent, as this effect was also seen in CL1-0 cells (another human cell line), (C) (D) Depletion of hHR23A upregulated Chk1 did not alter its mRNA expression.Supplemental Fig. 2(A) (B) Chk1 associates with hHR23A but not hHR23B. 26 RAD23A UNIPROT:P54725 decreases UNIPROT:O14757 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 We found that hHR23A overexpression dose-dependently decreased the levels of Chk1 in sh-HR23A cells (Fig. 3A).|||The induction of Chk1 protein levels by hHR23A knockdown was not cell-context-dependent, as this effect was also seen in CL1-0 cells (another human cell line) (Supplemental Fig. 1B), AGS and MKN-45 cells (Fig. 2C and D).|||The induction of Chk1 protein levels by hHR23A knockdown was not cell-context-dependent, as this effect was also seen in CL1-0 cells (another human cell line) (Supplemental Fig. 1B), AGS and MKN-45 cells (Fig. 2C and D).|||We show that hHR23A associates with Chk1 through the UBA domains, and that silencing of hHR23A increases the protein level and S347 phosphorylation of Chk1.|||In cells subjected to cisplatin-induced DNA damage, depletion of hHR23A further augmented the protein and phosphorylation levels of Chk1 (Fig. 6A and B), consistent with the notion that Chk1 functions in the DDR.|||DNA damaging agents reduce the interaction between hHR23A and Chk1 In cells subjected to cisplatin-induced DNA damage, depletion of hHR23A further augmented the protein and phosphorylation levels of Chk1 (Fig. 6A and B), consistent with the notion that Chk1 functions in the DDR.|||Knockdown of hHR23A enhances the protein level and stability of Chk1 The increase in Chk1 protein levels observed under hHR23A knockdown prompted us to question whether hHR23A regulates the protein expression and function of Chk1.|||(B) The induction of Chk1 protein levels by hHR23A knockdown was not cell-context-dependent, as this effect was also seen in CL1-0 cells (another human cell line), (C) (D) Depletion of hHR23A upregulated Chk1 did not alter its mRNA expression.Supplemental Fig. 2(A) (B) Chk1 associates with hHR23A but not hHR23B. 16 RAD23A UNIPROT:P54725 activates UNIPROT:P68036 Protein 2f658338-cc8d-11e5-aade-001a4ae51247 9153201 However, upon addition of bacterially expressed UbcH8 or UbcH7, the ubiquitination of HHR23A was enhanced (lanes 3and5). 16 RAD23A UNIPROT:P54725 activates UNIPROT:O14933 Protein 2f658338-cc8d-11e5-aade-001a4ae51247 9153201 However, upon addition of bacterially expressed UbcH8 or UbcH7, the ubiquitination of HHR23A was enhanced (lanes 3and5). 16 MESH:D002110 ubiquitinatesProtein RAD23A UNIPROT:P54725 Phenotype 41749470-cbf1-11e5-b0dd-001a4ae51247 10373495 In order to ascertain whether E6AP is involved in thein vivoubiquitination of HHR23A, wild-type and mutant forms of E6AP were transfected into mammalian cells, and their expression was confirmed by immunoblot analysis with anti-E6AP antisera (32).|||Effect of Wild-type and Mutant E6AP on HHR23A Ubiquitination in Vivo In order to ascertain whether E6AP is involved in thein vivoubiquitination of HHR23A, wild-type and mutant forms of E6AP were transfected into mammalian cells, and their expression was confirmed by immunoblot analysis with anti-E6AP antisera (32). 10 RAD23A UNIPROT:P54725 activates GO:0051320 Phenotype 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Conversely, ectopic expression of hHR23A rescued the accumulation of sh-HR23A cells in the S phase to control levels (Fig. 2E).|||Knockdown of hHR23A attenuates cell cycle progression at S phase To investigate the distinct roles of human Rad23A (hHR23A) and Rad23B (hHR23B), we previously established hHR23A- and hHR23B-knockdown cell lines using a lentiviral shRNA mediated approach in A549 cells[25]. 8 RAD23A UNIPROT:P54725 activates UNIPROT:P04637 Protein df64eb42-3aaa-11e8-87fd-001a4a160176 27771451 siRNA-mediated depletion of HR23A, HR23B, or both proteins stimulates rapid p53 turnover by the proteasome (Brignone et al., 2004; Kaur et al., 2007) and attenuates the p53-dependent apoptotic signal after exposure to genotoxic stress (Kaur et al., 2007). 8 RAD23A UNIPROT:P54725 activates GO:0051726 Phenotype 5bedf590-3511-11e8-a34b-001a4a160175 26296656 We reasoned that hHR23A might modulate a specific cell cycle regulator, leading to the delay in cell cycle progression. 8 RAD23A UNIPROT:P54725 activates GO:0006915 Phenotype d021c75a-2804-11f0-b759-0050569a791b 10.1016/s1016-8478(23)12897-4 Gaynor and Chen (2001) found that overexpression of hHR23A could induce solely apoptosis, and that mutation of its C-terminal ubiquitin associated (UBA) domain, a Vpr-binding domain, caused disruption of the apoptotic effects. 6 RAD23A UNIPROT:P54725 decreases UNIPROT:P04637 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "However, coexpression of increasing amounts of hHR23A caused a dose-dependent reduction in the observed levels of p53 and consequently decreased the levels of p21waf1(Fig. 5A)⇓.|||To test whether MDM2 and hHR23A could act cooperatively to inhibit p53-dependent transcription, we first evaluated the effects of overproduction of p300 (300-528) on MDM2-mediated inhibition of p53-dependent transcription.|||Compared with the expression of p300 segments alone, coexpression of hHR23A inhibited the p53-dependent transcription stimulated by p300 (1-140), p300 (1-340), and p300 (1-595) (Fig. 3B⇓,column 5 versus column 4,column 7 versus column 6, andcolumn 9 versus column 8).|||hHR23A, however, failed to inhibit p53-dependent transcription stimulated by p300 (300-528).|||To this end, our results demonstrated that by interacting with the p300/CBP C/H1 domain, overexpression of hHR23A inhibits p53 transcriptional activity, decreases the steady-state level of p53, and subsequently reduces the level of p21waf1.|||However, this explanation cannot account for why overexpression of hHR23A also causes a decrease in the level of p53." 4 RAD23A UNIPROT:P54725 activates UNIPROT:O14757 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 In contrast, overexpression of hHR23A enhanced the Ub level on Chk1 compared with that seen in sh-HR23A cells (Fig. 5B). 4 RAD23A UNIPROT:P54725 inhibits UNIPROT:P04637 Protein df64eb42-3aaa-11e8-87fd-001a4a160176 27771451 On the other hand, overexpression of hHR23A inhibits p53-mediated transactivation, and this inhibitory effect is abolished by co-expression of the transcriptional co-activator p300 (Zhu et al., 2001). 4 RAD23A UNIPROT:P54725 inhibits FPLX:Proteasome ProteinFamily aa71bbd8-338c-11e8-b868-001a4a160176 16860562 Extreme overexpression of either HR23A or HR23B in living cells is known to inhibit 26S proteasome-mediated breakdown (Ortolan et al., 2000; Raasi and Pickart, 2003). 4 RAD23A UNIPROT:P54725 increases FPLX:Ubiquitin ProteinFamily 5bedf590-3511-11e8-a34b-001a4a160175 26296656 In contrast, overexpression of hHR23A enhanced the Ub level on Chk1 compared with that seen in sh-HR23A cells (Fig. 5B). 4 RAD23A UNIPROT:P54725 increases UNIPROT:O14757 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 In this study, we observed that treatment with the deubiquitinase inhibitor Ub aldehyde increased the level of Chk1 poly-ubiquitination (Fig. 5C), and ectopic expression of hHR23A rescued the interaction of Chk1 and USP7 to control levels (Fig. 5D).|||DNA damaging agents reduce the interaction between hHR23A and Chk1 In cells subjected to cisplatin-induced DNA damage, depletion of hHR23A further augmented the protein and phosphorylation levels of Chk1 (Fig. 6A and B), consistent with the notion that Chk1 functions in the DDR. 4 RAD23A UNIPROT:P54725 inhibits UNIPROT:Q12933 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 As shown in (Fig. 4C), RAD23A overexpression caused a decrease of TRAF2 half-life, which indicates that RAD23A accelerates TRAF2 degradation.|||Furthermore, we found that RAD23A down-regulate TRAF2 by promoting its degradation through ubiquitin–proteasome system.|||RAD23A promotes ubiquitin-proteasome dependent TRAF2 degradation As an UbL-UBA domain containing protein, RAD23A was reported to promote substrate recognition by proteasome[24]. 4 RAD23A UNIPROT:P54725 inhibits UNIPROT:P04637 Protein c5184a12-cb27-11e5-8189-001a4ae51246 11340074 Moreover, overexpression of wild-type hHR23A inhibits the p53 transcriptional activity and results in a decreased steady-state protein level of cellular p53 (28).|||First, we have already established that hHR23A interacts with the CH1 region (aa 325–528) of p300/CBP to down-regulate p53 (28). 4 UNIPROT:Q17RS7 activates RAD23A UNIPROT:P54725 Protein cf0747ac-3403-11e8-9fbf-001a4a160176 26095601 Additionally, hHR23A, a gene related to DNA repair, was significantly up-regulated by 1 or 5 µM GEN treatment compared with the R group (P<0.05) (Fig. 7C). 4 FPLX:PKA increases RAD23A UNIPROT:P54725 ProteinFamily ddd423d6-ab97-11e6-9646-001a4ae51246 PMC4537845 Likewise, co-expression of WT UBE3A with increasing amounts of constitutively active PKA (PKA-CA) led to increased levels of HHR23A, while HHR23A levels remained low in cells expressing UBE3A-T485A and PKA-CA (Figures S4C and S4D). 4 MESH:D002945 increases RAD23A UNIPROT:P54725 Phenotype 5dc95a26-3534-11e8-a51f-001a4a160176 21742020 A time course analysis showed that 1μg/ml cisplatin time-dependently enhanced hHR23A and hHR23B protein expression (1.7-fold for hHR23A and hHR23B at 6-h treatment) (Fig. 1B) and then declined to basal level after 36h, indicating that a low dose of cisplatin transiently enhanced hHR23A and hHR23B protein levels.|||1A, cisplatin significantly induced hHR23A and hHR23B protein expression in a dose-dependent manner, with a 2-fold increase at 1μg/ml cisplatin treatment.|||Collectively, we speculated that the transient enhancement of hHR23A and hHR23B protein levels by cisplatin reflected post-transcriptional regulation. 3 RAD23A UNIPROT:P54725 inhibits UNIPROT:Q09472 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "Coexpression of hHR23A almost completely blocked the stimulatory effects of p300 (1-140) and p300 (1-340), but only decreased enhancement of the expression of the reporter from ∼3-fold by p300 (1-595) to 2-fold.|||Accordingly, the expression of hHR23A antagonizes the stimulatory effects of p300 on p53-dependent expression." 3 RAD23A UNIPROT:P54725 activates UNIPROT:P42737 Protein 251e75f0-bc44-11e5-9b9d-001a4ae51247 PMC3298473 Figure4Band4Cshowed that hHR23a increased the stability of BCA2 through the inhibition autoubiquitination.|||These observations were strengthened since hHR23a was also shown to increase the half-life of BCA2 over a 24 hours time period (Figure4D).|||Taken together, data presented here argues that hHR23a presence may be necessary to modulate and regulate BCA2 in a cancer setting. 2 RAD23A UNIPROT:P54725 increases UNIPROT:Q01094 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 We observed that endogenous E2F1 levels are increased by UVB radiation or by expression of hHR23A, and the presence of exogenous hHR23A further increased E2F1 levels in UVB-treated keratinocytes (Supplementary Figure S3). 2 RAD23A UNIPROT:P54725 activates UNIPROT:Q09472 Protein c5184a12-cb27-11e5-8189-001a4ae51246 11340074 A. Wani, unpublished observation.Therefore, a model is proposed to suggest that the hHR23A protein, through interaction with CH1 domain, enables the docking of proteasome onto p300 (Fig.7). 2 RAD23A UNIPROT:P54725 inhibits UNIPROT:P42739 Protein 7a97940a-bc3d-11e5-9b9d-001a4ae51247 PMC3864866 HHR23a UBA domains sequester polyubiquitin By using a previously established assay,30we determined that the UBA domains of hHR23a are highly efficient at sequestering the ubiquitin moieties of K48-linked and K63-linked tetraubiquitin. 2 RAD23A UNIPROT:P54725 activates UNIPROT:P04637 Protein 2f291a94-bc00-11e5-8abe-001a4ae51246 10.1016/j.bbrc.2005.07.067 Overexpression of hHR23A and B proteins has led to the accumulation of ubiquitinated p53 and blocked p53 proteasome degradation[17]. 2 RAD23A UNIPROT:P54725 inhibits UNIPROT:P04637 Protein 2f291a94-bc00-11e5-8abe-001a4ae51246 10.1016/j.bbrc.2005.07.067 hHR23A and B proteins downregulated the transactivating activity of p53 by interacting with the p300/cyclic AMP-responsive element binding (CREB)-binding protein[18]. 2 RAD23A UNIPROT:P54725 activates UNIPROT:P54725 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Knockdown of hHR23A attenuates cell cycle progression at S phase To investigate the distinct roles of human Rad23A (hHR23A) and Rad23B (hHR23B), we previously established hHR23A- and hHR23B-knockdown cell lines using a lentiviral shRNA mediated approach in A549 cells[25]. 2 RAD23A UNIPROT:P54725 ubiquitinatesProtein UNIPROT:P54725 Protein 2f658338-cc8d-11e5-aade-001a4ae51247 9153201 Finally, we demonstrate that the two E6AP-interacting E2s also functioned in conjunction with E6AP in the ubiquitination of HHR23A, an E6AP substrate (22).2The noninteracting E2s were unable to enhance the ubiquitination of HHR23A above background levels observed in wheat germ extracts. 2 RAD23A UNIPROT:P54725 inhibits UNIPROT:P54727 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 (B) The induction of Chk1 protein levels by hHR23A knockdown was not cell-context-dependent, as this effect was also seen in CL1-0 cells (another human cell line), (C) (D) Depletion of hHR23A upregulated Chk1 did not alter its mRNA expression.Supplemental Fig. 2(A) (B) Chk1 associates with hHR23A but not hHR23B. 2 RAD23A UNIPROT:P54725 activates UNIPROT:P54727 Protein 68c3080a-c85e-11ee-b346-0050569a791b 10.1016/j.bbagrm.2023.194925 However, RAD23A is less abundant compared to RAD23B and accordingly it has been observed that in mouse fibroblasts, RAD23A needs to be overexpressed to substitute RAD23B in XPC stabilization[41]. 2 RAD23A UNIPROT:P54725 activates UNIPROT:P54727 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Knockdown of hHR23A attenuates cell cycle progression at S phase To investigate the distinct roles of human Rad23A (hHR23A) and Rad23B (hHR23B), we previously established hHR23A- and hHR23B-knockdown cell lines using a lentiviral shRNA mediated approach in A549 cells[25]. 2 RAD23A UNIPROT:P54725 activates UNIPROT:Q01831 Protein 68c3080a-c85e-11ee-b346-0050569a791b 10.1016/j.bbagrm.2023.194925 However, RAD23A is less abundant compared to RAD23B and accordingly it has been observed that in mouse fibroblasts, RAD23A needs to be overexpressed to substitute RAD23B in XPC stabilization[41]. 2 RAD23A UNIPROT:P54725 activates GO:0007049 Phenotype 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Knockdown of hHR23A attenuates cell cycle progression at S phase To investigate the distinct roles of human Rad23A (hHR23A) and Rad23B (hHR23B), we previously established hHR23A- and hHR23B-knockdown cell lines using a lentiviral shRNA mediated approach in A549 cells[25]. 2 RAD23A UNIPROT:P54725 activates GO:0042769 Phenotype 5bedf590-3511-11e8-a34b-001a4a160175 26296656 DNA damaging agents reduce the interaction between hHR23A and Chk1 In cells subjected to cisplatin-induced DNA damage, depletion of hHR23A further augmented the protein and phosphorylation levels of Chk1 (Fig. 6A and B), consistent with the notion that Chk1 functions in the DDR. 2 RAD23A UNIPROT:P54725 inhibits GO:1903009 Phenotype 2f291a94-bc00-11e5-8abe-001a4ae51246 10.1016/j.bbrc.2005.07.067 Overexpression of hHR23A and B proteins has led to the accumulation of ubiquitinated p53 and blocked p53 proteasome degradation[17]. 2 RAD23A UNIPROT:P54725 inhibits GO:0006289 Phenotype 2f291a94-bc00-11e5-8abe-001a4ae51246 10.1016/j.bbrc.2005.07.067 This difference presumably contributes to the following findings: (1) knock down of hHR23A protein causes mild deficiency of NER and (2) knock down of hHR23B protein causes a substantial inhibition of NER activity. 2 RAD23A UNIPROT:P54725 activates FPLX:Proteasome ProteinFamily c5184a12-cb27-11e5-8189-001a4ae51246 11340074 A. Wani, unpublished observation.Therefore, a model is proposed to suggest that the hHR23A protein, through interaction with CH1 domain, enables the docking of proteasome onto p300 (Fig.7). 2 RAD23A UNIPROT:P54725 ubiquitinatesProtein UNIPROT:Q01094 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 The presence of either hHR23A or hHR23B substantially increased the abundance of ubiquitylated E2F1, irrespective of the differentiation status of the keratinocytes.|||Next, we determined the effect of proteasomal inhibition on modulation by hHR23A of polyubiquitylated E2F1. 2 RAD23A UNIPROT:P54725 decreases UNIPROT:Q09472 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "Additionally, coexpression of hHR23A was also found to cause a prominent decrease in the steady-state level of the p300 segment, p300 (300-528) (data not shown).|||Taken together, these results indicate that although the COOH-terminal 57 amino acid residues of hHR23A were enough to inhibit p53-independent and presumably p300/CBP-dependent transcription, this segment of hHR23A was not sufficient to influence the p53-dependent transcription of target genes." 2 RAD23A UNIPROT:P54725 inhibits UNIPROT:P04637 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "The data demonstrate that hHR23A can interact with p300/CBP to down-regulate p53 transcriptional activity and suggest that hHR23 proteins play a key role in cross-talk between the p53 pathway and DNA repair.|||As can be seen in Fig. 2A⇓, expression of p300 allowed a full recovery of the transcriptional activity of p53 inhibited by hHR23A (Fig. 2A⇓,columns 4–6), whereas expression of p300 alone increased the p53-dependent expression of the reporter to a maximum level." 2 UNIPROT:Q17RS7 increases RAD23A UNIPROT:P54725 Protein cf0747ac-3403-11e8-9fbf-001a4a160176 26095601 Besides HERP, hHR23A expression was also up-regulated by low-dose GEN 24 h after X-irradiation. 2 FPLX:Ubiquitin activates RAD23A UNIPROT:P54725 ProteinFamily 6085bc5a-bc3c-11e5-8abe-001a4ae51246 PMC4197637 Therefore, the exposed I44 patch of the proximal Ub, and valine-70 and leucine-73 residues of the distal Ub allow binding and positioning of the UBA2 domain of the proteasomal signaling protein hHR23A between the K48-linked chains [67]. 2 CHEBI:2150 decreases RAD23A UNIPROT:P54725 Chemical 3b454976-c8e7-11e5-a1fd-001a4ae51246 17848572 More interestingly, we observed that R1881, but not DHT, significantly decreased ABCG1 and RAD23A expression levels in control cells. 2 UNIPROT:P55072 activates RAD23A UNIPROT:P54725 Protein 445659f8-bc38-11e5-8abe-001a4ae51246 PMC3435318 hHR23A blocks the VCP/p97 stimulation of ataxin-3 ubiquitin hydrolase activity hHR23A and VCP/p97 are both involved in protein quality control through their connections with protein degradation and the ubiquitin-proteasome system[31]–[33]. 2 UNIPROT:P54725 activates RAD23A UNIPROT:P54725 Protein 5bedf590-3511-11e8-a34b-001a4a160175 26296656 Knockdown of hHR23A attenuates cell cycle progression at S phase To investigate the distinct roles of human Rad23A (hHR23A) and Rad23B (hHR23B), we previously established hHR23A- and hHR23B-knockdown cell lines using a lentiviral shRNA mediated approach in A549 cells[25]. 2 UNIPROT:P54725 ubiquitinatesProtein RAD23A UNIPROT:P54725 Protein 2f658338-cc8d-11e5-aade-001a4ae51247 9153201 Finally, we demonstrate that the two E6AP-interacting E2s also functioned in conjunction with E6AP in the ubiquitination of HHR23A, an E6AP substrate (22).2The noninteracting E2s were unable to enhance the ubiquitination of HHR23A above background levels observed in wheat germ extracts. 2 MESH:D015741 decreases RAD23A UNIPROT:P54725 Phenotype 3b454976-c8e7-11e5-a1fd-001a4ae51246 17848572 More interestingly, we observed that R1881, but not DHT, significantly decreased ABCG1 and RAD23A expression levels in control cells. 2 FPLX:E3:Ub:ligase activates RAD23A UNIPROT:P54725 ProteinFamily 83a1cf4a-ca00-11e5-b88f-001a4ae51247 15280365 Previous studies of the hHR23A protein showed that overexpression of the E3 ligase E6AP enhanced ubiquitin modification of hHR23A and that catalytically inactive (C833A) E6AP failed to promote ubiquitination of hHR23A (39). 2 UNIPROT:P51668 ubiquitinatesProtein RAD23A UNIPROT:P54725 Protein 917a3286-ca02-11e5-b88f-001a4ae51247 PMC1356336 The ability of submicromolar concentrations of UbcH5A to support the ubiquitylation of HHR23A suggests that UbcH5A binds efficiently to E6AP.|||Our finding that UbcH5A and E6AP can produce strong ubiquitylation of HHR23A contrasts with an earlier report that UbcH5A is an inefficient partner of E6AP (Kumaret al, 1997). 2 FPLX:E3:Ub:ligase ubiquitinatesProtein RAD23A UNIPROT:P54725 ProteinFamily 1168319c-7a08-11ee-add2-0050569a791b PMC10169902 We found that the overexpression of the E3 E6AP leads to increased ubiquitylation of RAD23A, especially when lysines were introduced into the lysine desert.|||Since human RAD23A was previously shown to be ubiquitylated by the E3 E6AP [35] (UBE3A), we tested if ubiquitylation of the lysine version of RAD23A was affected upon overexpression of E6AP. 1 RAD23A UNIPROT:P54725 phosphorylatesProtein UNIPROT:O14757 Protein a3a448e7-fb0b-11e9-b12d-001a4a160176 PMC6834284 It may be possible that RAD23A phosphorylates CHEK1 since its cousin, RAD17, is a known phosphorylator of CHEK1 [25]. 1 RAD23A UNIPROT:P54725 activates UNIPROT:O14757 Protein a3a448e7-fb0b-11e9-b12d-001a4a160176 PMC6834284 Thus the reduction of RAD23A with longevity could inhibit CHEK1 degredation and allow promotion of tumor growth and therapy resistance. 1 RAD23A UNIPROT:P54725 decreases UNIPROT:O14757 Protein a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Our group previously showed that hHR23A, but not hHR23B, is essential for the Chk1 turnover; and loss of hHR23A increases Chk1 protein expression while attenuating cell division and cell cycle progression[35]. 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q9BYX4 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Results and discussion Knockdown of RAD23A augments RIG-I/MDA5 mediated signaling activation RIG-I/MDA5 recognizes viral RNAs in cytoplasm and triggers signal transduction cascades that finally lead to the production of type I IFNs and proinflammatory cytokines to suppress viral replication and assembly. 1 RAD23A UNIPROT:P54725 ubiquitinatesProtein UNIPROT:P05928 Protein 9771fef4-ae93-11ec-a61d-0050569a1f61 PMCPMC8617076 Interestingly, in contrast to other DNA repair proteins that are targeted by Vpr, hHR23A does not become ubiquitinated in a Vpr-dependent manner and is not depleted in cells expressing HIV-1 Vpr , suggesting that hHR23A is not a Vpr-recruited CRL4 E3 substrate and that the Vpr–hHR23A complex likely presents a distinct architecture.Here, we report the high-resolution NMR structure of Vpr, complexed with the C-terminal half of hHR23A, containing the XPCB and UBA2 domains, and a 2. 1 RAD23A UNIPROT:P54725 activates UNIPROT:P05928 Protein 74ea177c-bc37-11e5-9b9d-001a4ae51247 PMC2894085 Most significantly (p<0.05), depletion of hHR23A reduced replication of the Vpr(+) virus by over 65% (Fig. 3Aii), whereas no effect was observed on replication of the Vpr(−) HIV-1. 1 RAD23A UNIPROT:P54725 deubiquitinatesProtein UNIPROT:P42737 Protein 251e75f0-bc44-11e5-9b9d-001a4ae51247 PMC3298473 hHR23a inhibits BCA2 autoubiquitination activity and stabilizes BCA2 In its role as an ubiquitin receptor, hHR23a is known to prevent the formation of polyubiquitin chains, and thus inhibits degradation of target proteins [22,23]. 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q12933 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 We further found that the effect of RAD23A on TRAF2 protein level was blocked by proteasome inhibitor MG132 (Fig. 4D), suggesting that the ubiquitin-proteasome pathway is required for RAD23A mediated downregulation of TRAF2. 1 RAD23A UNIPROT:P54725 ubiquitinatesProtein UNIPROT:Q12933 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Our data implicated that RAD23A might increase TRAF2 ubiquitination by inhibiting its deubiquitylation. 1 RAD23A UNIPROT:P54725 decreases UNIPROT:O00429 Protein 55ba2e1a-bc22-11e5-9b9d-001a4ae51247 PMC3391904 In this tumor, DRP1 is overexpressed, but is sequestered in the nucleus by hHR23A, so avoiding its localization on mitochondria and conferring resistance to cisplatin [56]. 1 RAD23A UNIPROT:P54725 increases UNIPROT:O00429 Protein a9a75fd4-bc36-11e5-ac4e-001a4ae51246 PMC3041770 Silencing of hHR23A, a nucleotide excision repair (NER) enzyme, decreases the nuclear DRP1 level and cisplatin resistance in lung adenocarcinoma cells[31]. 1 RAD23A UNIPROT:P54725 activates UNIPROT:P54252 Protein 445659f8-bc38-11e5-8abe-001a4ae51246 PMC3435318 Here, we sought to investigate whether VCP/p97 or hHR23A, both of which are involved in protein degradation pathways, also enhanced ataxin-3 activity through direct interactions. 1 RAD23A UNIPROT:P54725 activates UNIPROT:P54252 Protein 6ea9fadc-374e-11e8-a51f-001a4a160176 28928079 HHR23A is found in nuclear inclusions of expanded polyQ Atx3 (Doss-Pepe et al., 2003; Wang et al., 2000) and was shown to prevent Atx3 proteasomal degradation (Blount et al., 2014). 1 RAD23A UNIPROT:P54725 inhibits UNIPROT:O15265 Protein ab1bc7b6-bbf9-11e5-8abe-001a4ae51246 PMC4237202 It would be of interest to determine if Rad23A inhibits the degradation of ataxin-7, similar to its effect on ataxin-3. 1 RAD23A UNIPROT:P54725 deubiquitinatesProtein UNIPROT:Q01094 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 These observations suggest that hHR23A may preferentially modulate the degradation of poly-ubiquitylated E2F1, although mono-ubiquitin modification of the latter on one or more Lys residues likely also takes place in keratinocytes, without precluding its proteasomal degradation. 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q01094 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 Reciprocally, hHR23A is able to reduce E2F1 turnover in undifferentiated keratinocytes. 1 RAD23A UNIPROT:P54725 decreases UNIPROT:Q92793 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "Taken together, these results indicate that although the COOH-terminal 57 amino acid residues of hHR23A were enough to inhibit p53-independent and presumably p300/CBP-dependent transcription, this segment of hHR23A was not sufficient to influence the p53-dependent transcription of target genes." 1 RAD23A UNIPROT:P54725 inhibits UNIPROT:P42739 Protein 0f71f906-bbfa-11e5-9b9d-001a4ae51247 PMC2748877 hHR23a is sandwiched between the ubiquitin subunits of K48-linked diubiquitin (Varadan et al., 2005), effectively sequesters subunits of polyubiquitin up through octaubiquitin (Kang et al., 2007), and protects ubiquitin chains from deubiquitination (Raasi and Pickart, 2003; Verma et al., 2004). 1 RAD23A UNIPROT:P54725 increases UNIPROT:P04637 Protein 80d76aa4-cb27-11e5-b419-001a4ae51246 11196199 "Compared with the expression of p300 segments alone, coexpression of hHR23A inhibited the p53-dependent transcription stimulated by p300 (1-140), p300 (1-340), and p300 (1-595) (Fig. 3B⇓,column 5 versus column 4,column 7 versus column 6, andcolumn 9 versus column 8)." 1 RAD23A UNIPROT:P54725 decreases UNIPROT:P04637 Protein 5dc95a26-3534-11e8-a51f-001a4a160176 21742020 In contrast, overexpression of hHR23A downregulates steady-state p53 protein levels and transcriptional activity through interactions with p300/CBP (Zhu et al., 2001). 1 RAD23A UNIPROT:P54725 inhibits UNIPROT:P04637 Protein 1ad13a88-c8ec-11e5-8b47-001a4ae51246 PMC2696100 In one investigation siRNA-mediated knockdown of hHR23 A and B caused accumulation of p53, consistent with a role in transport of p53 to the proteasome (77). 1 RAD23A UNIPROT:P54725 ubiquitinatesProtein UNIPROT:P04637 Protein f4855616-c472-11e5-9da3-001a4ae51247 PMC2775171 Both HR23A and ubiquilin 1 TUBEs efficiently purified ubiquitylated p53 (Fig 5B). 1 RAD23A UNIPROT:P54725 activates UNIPROT:O00421 Protein 7fafdb80-bbea-11e5-8abe-001a4ae51246 10.1016/j.dnarep.2011.05.003 In non-irradiated cells, a decrease only in RAD23A gene expression reduced HCR, highlighting the different roles of the human RAD23 genes in repair. 1 RAD23A UNIPROT:P54725 activates UNIPROT:O75976 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 Thus, the joint expression of E2F1 and hHR23A accelerates CPD removal and DNA repair following UVB damage. 1 RAD23A UNIPROT:P54725 increases UNIPROT:P54725 Protein ab1bc7b6-bbf9-11e5-8abe-001a4ae51246 PMC4237202 Knockdown of endogenous Rad23A or Rad23B through different, non-overlapping siRNA constructs noticeably decreases levels of Rad23A or Rad23B protein. 1 RAD23A UNIPROT:P54725 inhibits UNIPROT:P54725 Protein 41749470-cbf1-11e5-b0dd-001a4ae51247 10373495 The decrease in HHR23A protein levels was completely blocked by transient expression of dominant negative E6AP, providing direct evidence for the involvement of E6AP in the cell cycle-dependent degradation of HHR23A. 1 RAD23A UNIPROT:P54725 increases UNIPROT:P54727 Protein ab1bc7b6-bbf9-11e5-8abe-001a4ae51246 PMC4237202 Knockdown of endogenous Rad23A or Rad23B through different, non-overlapping siRNA constructs noticeably decreases levels of Rad23A or Rad23B protein. 1 RAD23A UNIPROT:P54725 inhibits UNIPROT:P54727 Protein f65bce5a-ca00-11e5-ab20-001a4ae51246 15885096 This may result from the altered amino acid sequence (P331–V332 in hHR23B vs. P287–P288 in hHR23A), because the consecutive prolines of XPCB–hHR23A restrict the direction of the C-terminus in a different way to the Pro–Val sequence in hHR23B. 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q01831 Protein b0ce5a9c-bc45-11e5-8abe-001a4ae51246 10.1016/j.mrfmmm.2009.08.004 As far as their GG-NER functions are concerned, the two RAD23 homologues are functionally interchangeable, because overexpression of RAD23A can compensate for lack of RAD23B and thereby fully restores both XPC protein and GG-NER activities[59]. 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q01831 Protein b7b2c446-cbf0-11e5-b0dd-001a4ae51247 10197977 hHR23A can substitute for hHR23B in binding and stimulating XPC in vitro, suggesting some functional redundancy (Sugasawa et al. 1997). 1 RAD23A UNIPROT:P54725 activates UNIPROT:Q01831 Protein 7fafdb80-bbea-11e5-8abe-001a4ae51246 10.1016/j.dnarep.2011.05.003 Taken together, these results indicate that RAD23A and RAD23B proteins contribute to stabilize the XPC protein and lend further support to the hypothesis of Okuda et al. in which the interaction with RAD23B may stabilize the XPC protein pool in order to have a longer activation of the NER function[47]. 1 RAD23A UNIPROT:P54725 decreases UNIPROT:O95786 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 RAD23A negatively regulates RIG-I/MDA5-mediated expression of proinflammatory cytokines and IFNs. 1 RAD23A UNIPROT:P54725 activates UNIPROT:O95786 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Results and discussion Knockdown of RAD23A augments RIG-I/MDA5 mediated signaling activation RIG-I/MDA5 recognizes viral RNAs in cytoplasm and triggers signal transduction cascades that finally lead to the production of type I IFNs and proinflammatory cytokines to suppress viral replication and assembly. 1 RAD23A UNIPROT:P54725 activates UNIPROT:P01574 Protein 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Considering TRAF2 also plays essential roles in NF-κB activation in signalings such as TNFR and TLRs, we also tested the effect of RAD23A on TLR3 mediated NF-κB and IFN-β activation, and found that RAD23A exhibited similar function (data not shown). 1 RAD23A UNIPROT:P54725 activates GO:0044349 Phenotype 88e869b0-c469-11e5-9da3-001a4ae51247 PMC3830595 [25] Clinical studies have shown that trained cyclist[27] and marathon runners[26] exhibit upregulation of DNA excision repair enzymes NESP and RAD23A[27] as well as OGG1. 1 RAD23A UNIPROT:P54725 activates GO:0010506 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 hHR23A interacts with autophagic proteins To assess the potential hHR23A-mediated regulation of autophagy, we immunoprecipitated endogenous hHR23A and immunoblotted the isolated immunocomplexes using antibodies against LC3 and Beclin 1.Fig. 1 RAD23A UNIPROT:P54725 inhibits MESH:D009369 Phenotype a3a448e7-fb0b-11e9-b12d-001a4a160176 PMC6834284 Thus the reduction of RAD23A with longevity could inhibit CHEK1 degredation and allow promotion of tumor growth and therapy resistance. 1 RAD23A UNIPROT:P54725 inhibits GO:0006508 Phenotype 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Some studies suggested that RAD23A inhibits ubiquitination and ubiquitin dependent degradation, whereas other studies demonstrated that RAD23A could promote protein degradation by targeting proteins to the proteasome[24,25]. 1 RAD23A UNIPROT:P54725 inhibits GO:0006508 Phenotype b01afb4a-bbd3-11e5-8abe-001a4ae51246 PMC2535603 HHR23A interacts with 26S proteasome through its N-terminal UBL domain to promote protein degradation [26-28]. 1 RAD23A UNIPROT:P54725 activates GO:0006508 Phenotype 74ea177c-bc37-11e5-9b9d-001a4ae51247 PMC2894085 However, it is currently unknown whether the role of hHR23A in promoting proteolysis is universal to all poly-Ub proteins or is specific to a subset of functionally relevant target proteins. 1 RAD23A UNIPROT:P54725 inhibits GO:0039694 Phenotype 74ea177c-bc37-11e5-9b9d-001a4ae51247 PMC2894085 Most significantly, down-regulation of hHR23A reduces viral replication in non-dividing MAGI cells and macrophages, suggesting that hHR23A-mediated interaction of Vpr with proteasome plays an important role in viral replication in these non-dividing cells. 1 RAD23A UNIPROT:P54725 activates GO:0007049 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Our group previously showed that hHR23A, but not hHR23B, is essential for the Chk1 turnover; and loss of hHR23A increases Chk1 protein expression while attenuating cell division and cell cycle progression[35]. 1 RAD23A UNIPROT:P54725 inhibits GO:0007049 Phenotype 41749470-cbf1-11e5-b0dd-001a4ae51247 10373495 The decrease in HHR23A protein levels was completely blocked by transient expression of dominant negative E6AP, providing direct evidence for the involvement of E6AP in the cell cycle-dependent degradation of HHR23A. 1 RAD23A UNIPROT:P54725 activates GO:0006281 Phenotype 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 Thus, the joint expression of E2F1 and hHR23A accelerates CPD removal and DNA repair following UVB damage. 1 RAD23A UNIPROT:P54725 activates GO:0070911 Phenotype b0ce5a9c-bc45-11e5-8abe-001a4ae51246 10.1016/j.mrfmmm.2009.08.004 As far as their GG-NER functions are concerned, the two RAD23 homologues are functionally interchangeable, because overexpression of RAD23A can compensate for lack of RAD23B and thereby fully restores both XPC protein and GG-NER activities[59]. 1 RAD23A UNIPROT:P54725 activates MESH:D004298 Phenotype 5d84daf4-acfc-11ee-a9a2-0050569a1f61 10.1007/s00338-020-01952-4 Catabolism UV excision repair proteins (Rad23a) had increased DA at day 14 and then decreased differential abundance at days 21 and 31–33 (Fig.3). 1 RAD23A UNIPROT:P54725 activates MESH:D004249 Phenotype 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 Significantly, when E2F1 and hHR23A were co-expressed, both proteins concentrated in CPD-positive regions, indicating that E2F1 promotes enrichment of hHR23A at sites of UV-induced DNA damage in primary keratinocytes (Figure7A). 1 RAD23A UNIPROT:P54725 activates GO:0051301 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Our group previously showed that hHR23A, but not hHR23B, is essential for the Chk1 turnover; and loss of hHR23A increases Chk1 protein expression while attenuating cell division and cell cycle progression[35]. 1 RAD23A UNIPROT:P54725 activates MESH:D002945 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Moreover, ectopic expression of hHR23A in sh-HR23A cells rescued the sensitivity of these cells to cisplatin (Fig. 1C) and oxaliplatin (Fig. 1D). 1 RAD23A UNIPROT:P54725 inhibits GO:1903009 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Given that hHR23A functions in ubiquitin-mediated proteasome degradation, we believe that hHR23A may regulate autophagy process by interacting with poly-ubiquitin conjugates on autophagy-related proteins, thereby acting as an ubiquitin chain carrier. 1 RAD23A UNIPROT:P54725 activates GO:0006289 Phenotype d15f8730-bc05-11e5-8abe-001a4ae51246 10.1016/j.yexcr.2008.10.027 The 19S regulatory particle is thought to have chaperone-like functions vital to nuclear excision repair (NER) mediated by the action of RAD23A. 1 RAD23A UNIPROT:P54725 activates MESH:D000077150 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Moreover, ectopic expression of hHR23A in sh-HR23A cells rescued the sensitivity of these cells to cisplatin (Fig. 1C) and oxaliplatin (Fig. 1D). 1 RAD23A UNIPROT:P54725 activates GO:0016049 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 Ectopic expression of His-tagged hHR23A increased cell growth rate compared to sh-HR23A cells. 1 RAD23A UNIPROT:P54725 activates GO:0006914 Phenotype a3d1a7e8-3387-11e8-87fd-001a4a160176 27613096 We further report that hHR23A modulates autophagy by interacting with autophagy-related proteins, such as LC3 and Beclin 1. 1 RAD23A UNIPROT:P54725 inhibits IP:IPR006155 ProteinFamily 3d89595a-bc37-11e5-ac4e-001a4ae51246 PMC2928749 However, full-length hHR23A does reduce the ability of the isolated Josephin domain to cleave polyUb chains (Figs. 1 RAD23A UNIPROT:P54725 increases FPLX:Proteasome ProteinFamily 3a018b3c-bbfa-11e5-8abe-001a4ae51246 PMC3038635 Addition of hHR23A at 4°C stimulated the maximal amount of proteasome binding to ubiquitinated E6AP with a Kaof approximately 16 nM (data not shown). 1 RAD23A UNIPROT:P54725 inhibits FPLX:Proteasome ProteinFamily 690a667e-bbff-11e5-9b9d-001a4ae51247 10.1016/j.bbrc.2013.01.059 Furthermore, we found that RAD23A down-regulate TRAF2 by promoting its degradation through ubiquitin–proteasome system. 1 RAD23A UNIPROT:P54725 activates FPLX:ERK ProteinFamily 5dc95a26-3534-11e8-a51f-001a4a160176 21742020 Conversely, ectopic expression of constitutively active MEK1 and MEK2 increased hHR23A and hHR23B protein levels (Fig. 5C), confirming that cisplatin upregulates hHR23 protein expression, at least in part, through MEK–ERK1/2 signaling. 1 RAD23A UNIPROT:P54725 activates IP:IPR004806 ProteinFamily 7fafdb80-bbea-11e5-8abe-001a4ae51246 10.1016/j.dnarep.2011.05.003 In non-irradiated cells, a decrease only in RAD23A gene expression reduced HCR, highlighting the different roles of the human RAD23 genes in repair. 1 RAD23A UNIPROT:P54725 inhibits PF:PF00627 ProteinFamily 2366370c-c6b3-11ee-9133-0050569a1f61 10.1016/j.jmb.2023.168353 Indeed, mutation of the RAD23A UBA domains disrupts binding to each polymerase, suggesting each interacts with the same UBA surfaces. 1 UNIPROT:Q9UMX0 increases RAD23A UNIPROT:P54725 Protein 1168319c-7a08-11ee-add2-0050569a791b PMC10169902 Quantification of the fluorescent signals by flow cytometry (Fig. S6) confirmed the results based on the blotting and microscopy (Fig.6), but also revealed slightly increased levels of the RAD23A variants as compared to wild type, as well as slightly increased levels of the R → Q variants of UBQLN1 and BAG6 as compared to wild type (Fig. S6). 1 FPLX:Ubiquitin activates RAD23A UNIPROT:P54725 ProteinFamily 914db5ce-ca5e-11e5-9bd2-001a4ae51247 PMC240680 "We predict that ubiquitin binds each of the UBA domains in a manner similar to that of the UBL domain and also causes hHR23a to adopt an opened conformation." 1 FPLX:Ubiquitin inhibits RAD23A UNIPROT:P54725 ProteinFamily e7ef6024-cb28-11e5-9aa0-001a4ae51247 PMC312714 "Polyubiquitination and ubiquitin-dependent degradation of HHR23A protein has been observed during the S phase of the cell cycle in human cells infected with human papilloma virus (Kumar et al. 1999)." 1 UNIPROT:P05928 activates RAD23A UNIPROT:P54725 Protein 74ea177c-bc37-11e5-9b9d-001a4ae51247 PMC2894085 Thus the expression of Vpr appeared to promote hHR23A-mediated protein poly-Ub (Fig. 2D, lanes 2, 4 and 6). 1 UNIPROT:P05928 activates RAD23A UNIPROT:P54725 Protein 61c565f6-c467-11e5-9da3-001a4ae51247 PMC3908392 On the other hand, given the ability of the UBA2 (and UBA1) domain to interact with poly-ubiquitinated proteins, complex formation between Vpr and the UBA2 and XPCB domains of hHR23A may modulate the role of hHR23A as a shuttle factor (61–64). 1 UNIPROT:Q4L180 activates RAD23A UNIPROT:P54725 Protein d8ca3608-c47c-11e5-9cc6-001a4ae51246 PMC3173115 FILIP-1L expression in cells leads to an increase in Hsf1 ubiquitination and its recruitment to the complexes with the UBA domain of hHR23A protein. 1 UNIPROT:P51451 ubiquitinatesProtein RAD23A UNIPROT:P54725 Protein 2aeed1d6-ca00-11e5-ab20-001a4ae51246 PMC522240 E6AP—which has been shown in noninfected cells to ubiquitinate HHR23A, a human Rad23 homologue (28), Mcm7 (27), and Blk, a Src family member (34)—is recruited by E6 to new substrates. 1 UNIPROT:Q01094 activates RAD23A UNIPROT:P54725 Protein 893a45d4-3799-11e6-aaca-001a4ae51246 PMC5041980 The ability of E2F1 to modulate hHR23A subcellular localization is further emphasized by the observation that both E2F1 and hHR23A remain cytoplasmic as a result of differentiation in keratinocytes. 1 UNIPROT:Q02750 increases RAD23A UNIPROT:P54725 Protein 5dc95a26-3534-11e8-a51f-001a4a160176 21742020 Conversely, ectopic expression of constitutively active MEK1 and MEK2 increased hHR23A and hHR23B protein levels (Fig. 5C), confirming that cisplatin upregulates hHR23 protein expression, at least in part, through MEK–ERK1/2 signaling. 1 UNIPROT:Q00987 activates RAD23A UNIPROT:P54725 Protein 12ef2cd2-bc51-11e5-9b9d-001a4ae51247 PMC4051618 There is evidence both for and against a role of the UBA/UBL adaptor proteins hHR23A and hHR23B in the degradation of p53.40,41,42,43Mdm2 can promote the ubiquitin-independent association of p53 with the proteasome.46There may be handover of ubiquitinated p53 from Mdm2 to S5a. 1 UNIPROT:P54725 inhibits RAD23A UNIPROT:P54725 Protein 41749470-cbf1-11e5-b0dd-001a4ae51247 10373495 The decrease in HHR23A protein levels was completely blocked by transient expression of dominant negative E6AP, providing direct evidence for the involvement of E6AP in the cell cycle-dependent degradation of HHR23A. 1 UNIPROT:P54725 increases RAD23A UNIPROT:P54725 Protein ab1bc7b6-bbf9-11e5-8abe-001a4ae51246 PMC4237202 Knockdown of endogenous Rad23A or Rad23B through different, non-overlapping siRNA constructs noticeably decreases levels of Rad23A or Rad23B protein. 1 UNIPROT:P54727 increases RAD23A UNIPROT:P54725 Protein ab1bc7b6-bbf9-11e5-8abe-001a4ae51246 PMC4237202 Knockdown of endogenous Rad23A or Rad23B through different, non-overlapping siRNA constructs noticeably decreases levels of Rad23A or Rad23B protein. 1 UNIPROT:P36507 increases RAD23A UNIPROT:P54725 Protein 5dc95a26-3534-11e8-a51f-001a4a160176 21742020 Conversely, ectopic expression of constitutively active MEK1 and MEK2 increased hHR23A and hHR23B protein levels (Fig. 5C), confirming that cisplatin upregulates hHR23 protein expression, at least in part, through MEK–ERK1/2 signaling. 1 UNIPROT:P03126 inhibits RAD23A UNIPROT:P54725 Protein fc307510-c8de-11e5-9cb8-001a4ae51247 16951183 "E6AP has also been shown to mediate the E6-dependent proteasomal degradation of hScrib and E6TP1 (41,42) and the E6-independent proteasomal degradation of Blk and HHR23A (43,44)." 1 UNIPROT:P05019 activates RAD23A UNIPROT:P54725 Protein eab3fce0-bc4e-11e5-9b9d-001a4ae51247 10.1016/j.mce.2010.10.009 qPCR Validation of the microarray data was conducted by performing qPCR on three genes that were upregulated by IGF1 (NFATC3,PPIP5K2, andTGFB2), three genes downregulated by IGF1 (RAD23A,H1FOO, andFADS6) and onGAPDH, which was used as a housekeeping gene. 1 UNIPROT:P05019 inhibits RAD23A UNIPROT:P54725 Protein eab3fce0-bc4e-11e5-9b9d-001a4ae51247 10.1016/j.mce.2010.10.009 qPCR Validation of the microarray data was conducted by performing qPCR on three genes that were upregulated by IGF1 (NFATC3,PPIP5K2, andTGFB2), three genes downregulated by IGF1 (RAD23A,H1FOO, andFADS6) and onGAPDH, which was used as a housekeeping gene. 1 UNIPROT:P33993 ubiquitinatesProtein RAD23A UNIPROT:P54725 Protein 2aeed1d6-ca00-11e5-ab20-001a4ae51246 PMC522240 E6AP—which has been shown in noninfected cells to ubiquitinate HHR23A, a human Rad23 homologue (28), Mcm7 (27), and Blk, a Src family member (34)—is recruited by E6 to new substrates. 1 IP:IPR004806 ubiquitinatesProtein RAD23A UNIPROT:P54725 ProteinFamily 2aeed1d6-ca00-11e5-ab20-001a4ae51246 PMC522240 E6AP—which has been shown in noninfected cells to ubiquitinate HHR23A, a human Rad23 homologue (28), Mcm7 (27), and Blk, a Src family member (34)—is recruited by E6 to new substrates. 1 UNIPROT:P46379 increases RAD23A UNIPROT:P54725 Protein 1168319c-7a08-11ee-add2-0050569a791b PMC10169902 Quantification of the fluorescent signals by flow cytometry (Fig. S6) confirmed the results based on the blotting and microscopy (Fig.6), but also revealed slightly increased levels of the RAD23A variants as compared to wild type, as well as slightly increased levels of the R → Q variants of UBQLN1 and BAG6 as compared to wild type (Fig. S6).