count source_label source_id relationship target_label target_id entity_type solr_id publication_id sentences 8 ANKLE1 UNIPROT:Q8NAG6 activates GO:0000422 Phenotype 537caebc-0550-11f0-bb39-0050569a791b 10.1016/j.bbamcr.2024.119752 Similarly, the ectopic overexpression of ANKLE1, responsible for cleaving the branched DNA, explicitly targeting the D-loop of mtDNA, led to the induction of mitophagy, consequently facilitating EMT in breast cancer. 8 ANKLE1 UNIPROT:Q8NAG6 activates GO:0006915 Phenotype 537caebc-0550-11f0-bb39-0050569a791b 10.1016/j.bbamcr.2024.119752 Moreover, ANKLE1 initiated genetic instability through the cleavage of nuclear DNA without inducing apoptosis [93]. 8 ANKLE1 UNIPROT:Q8NAG6 activates GO:0001837 Phenotype 537caebc-0550-11f0-bb39-0050569a791b 10.1016/j.bbamcr.2024.119752 Similarly, the ectopic overexpression of ANKLE1, responsible for cleaving the branched DNA, explicitly targeting the D-loop of mtDNA, led to the induction of mitophagy, consequently facilitating EMT in breast cancer. 4 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q12888 Protein 7e45fa48-c6a1-11ee-b346-0050569a791b 10.1016/j.tig.2023.10.004 ANKLE1 processes chromatin bridges induced by DNA-damaging agents to prevent the formation of G1-specific 53BP1 nuclear bodies, which represent DNA lesions. 4 ANKLE1 UNIPROT:Q8NAG6 activates UNIPROT:Q12888 Protein 7e45fa48-c6a1-11ee-b346-0050569a791b 10.1016/j.tig.2023.10.004 ANKLE1 processes chromatin bridges induced by DNA-damaging agents to prevent the formation of G1-specific 53BP1 nuclear bodies, which represent DNA lesions. 4 ANKLE1 UNIPROT:Q8NAG6 activates GO:0007059 Phenotype 7e45fa48-c6a1-11ee-b346-0050569a791b 10.1016/j.tig.2023.10.004 elegansembryos, LEM-3 processes chromatin bridges induced by incomplete DNA replication, unresolved recombination intermediates, and defective chromosome condensation to ensure chromosome segregation and cytokinesis [52]. 4 FPLX:RPA inhibits ANKLE1 UNIPROT:Q8NAG6 ProteinFamily 7e45fa48-c6a1-11ee-b346-0050569a791b 10.1016/j.tig.2023.10.004 This hypothesis is supported by the observation thatANKLE1deletion partially reduced the amount of TREX1-dependent RPA-coated ssDNA in chromatin bridges, suggesting ANKLE1 as one of the priming nucleases for TREX1 [35]. 4 UNIPROT:Q12476 phosphorylatesProtein ANKLE1 UNIPROT:Q8NAG6 Protein 7e45fa48-c6a1-11ee-b346-0050569a791b 10.1016/j.tig.2023.10.004 LEM-3 is phosphorylated at S192 and S194 by AIR-2/Aurora B kinase, which is required for the midbody localization of LEM-3 [52]. 2 ANKLE1 UNIPROT:Q8NAG6 activates UNIPROT:P55042 Protein 86a37944-c485-11e5-a92e-001a4ae51246 PMC4335191 elegansortholog of mammalian Ankle1,lem-3, causes a radiation hypersensitivity (Rad) phenotype (Christina Dittrich and Michael Hengartner, University of Zurich; personal communication). 2 ANKLE1 UNIPROT:Q8NAG6 activates GO:0002524 Phenotype 86a37944-c485-11e5-a92e-001a4ae51246 PMC4335191 elegansortholog of mammalian Ankle1,lem-3, causes a radiation hypersensitivity (Rad) phenotype (Christina Dittrich and Michael Hengartner, University of Zurich; personal communication). 2 ANKLE1 UNIPROT:Q8NAG6 activates MESH:D004249 Phenotype 86a37944-c485-11e5-a92e-001a4ae51246 PMC4335191 In order to find out whether Ankle1-mediated DNA cleavage activates DNA damage signaling, we analyzed the expression and localization of components of the DNA damage response pathway in GFP–Ankle1-expressing cells before and after Leptomycin treatment. 2 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q5ZXN6 Protein 16daab26-bbdb-11e5-9b9d-001a4ae51247 PMC4322725 Like AMPylation, phosphocholination is a reversible PTM.Tan et al. (2011)recently identified the Dot/Icm effector Lem3 as a protein capable to rescue the growth of yeast transformed by AnkX expression vector, which suggested that Lem3 was able to antagonize the activity of phosphocholination of AnkX.|||Indeed,in vitroassays demonstrated that Lem3 reverses the AnkX-dependent phosphocholination of Rab1 by removing the phosphocholine moiety from Rab1 (Tan et al., 2011;Table1). 2 ANKLE1 UNIPROT:Q8NAG6 activates GO:0008219 Phenotype 58fff77a-2a6e-11e6-9884-001a4ae51246 PMC4888674 However, as long-term expression of wild-type Ankle1 causes cell death (data not shown), we generated a stable U2OS cell line ectopically expressing an endonuclease-defective, GFP-tagged version of Ankle1 to address Ankle1 dynamics during the cell cycle and upon induction of DNA damage.|||This hypothesis is supported by previous findings showing that leptomycin B-mediated accumulation of Ankle1 in the nucleus causes DNA cleavage and cell death [22]. 2 CHEBI:16974 inhibits ANKLE1 UNIPROT:Q8NAG6 Chemical cbf41d20-c97a-11ee-b346-0050569a791b PMC10015115 Besides, both Lem1 and Lem3 proteins could be depleted by indole-3-acetic acid (IAA, auxin) in respective mutants, as shown by the disappearance of ∼130-kDa and ∼57-kDa bands in immunoblots, respectively (Fig. 3D).|||Immunofluorescence assay also confirmed the IAA-induced knockdown of Lem1 and Lem3 (Fig. 3E). 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q29ST3 Protein 0d7a1532-c47b-11e5-91a7-001a4ae51247 PMC3248503 Lem3 was unable to suppress the yeast toxicity of SidM (Fig. S4) (22), suggesting its specificity toward the activity of AnkX. 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:P34096 Protein 16daab26-bbdb-11e5-9b9d-001a4ae51247 PMC4322725 Indeed,in vitroassays demonstrated that Lem3 reverses the AnkX-dependent phosphocholination of Rab1 by removing the phosphocholine moiety from Rab1 (Tan et al., 2011;Table1). 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q5ZXN6 Protein ab9f4710-3b42-11e8-9fbf-001a4a160176 PMC6016468 pneumophilaeffector AnkX catalyzes the posttranslational addition of a PC moiety to Rab1A to prevent binding of host effectors and deactivation by GAPs (43), whereas Lem3 reverses the activity of AnkX by removal of phosphocholine from the switch II loop of Rab1A, which renders the protein susceptible to deactivation by GAPs (44). 1 ANKLE1 UNIPROT:Q8NAG6 activates UNIPROT:Q5ZXN6 Protein 11bbd972-bc48-11e5-8abe-001a4ae51246 PMC3558624 Furthermore,L. pneumophilais able to directly regulate AnkX activity by Lem3, which is a de-phosphorylcholinase that reverses the effects of AnkX on Rab1[17]. 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q5ZXN6 Protein 0d7a1532-c47b-11e5-91a7-001a4ae51247 PMC3248503 Together, these data suggest that Lem3 functions to antagonize the activity of AnkX. 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:Q5ZXN6 Protein 67c28cec-5c5d-11e7-bcb7-001a4ae51246 PMC5116710 "Ultimately, SidD antagonizes SidM function with a deAMPylation activity, and Lem3 reverses the effect of AnkX, thus allowing Rab1B to be inactivated by LepB, a GTPase activation protein (GAP) (89,91,92,103)." 1 ANKLE1 UNIPROT:Q8NAG6 activates UNIPROT:P14164 Protein 475f5406-bc38-11e5-8abe-001a4ae51246 PMC3285610 Our data suggest a model in which LEM-3 is required to support BAF-1 function. 1 ANKLE1 UNIPROT:Q8NAG6 inhibits UNIPROT:O00592 Protein 0d7a1532-c47b-11e5-91a7-001a4ae51247 PMC3248503 "Similar kinetics of Lem3-mediated PC moiety loss from PC-GST-Rab1 were observed when the samples were analyzed by mass spectrometry, in which the relative abundance of PCylated peptide -T72ITSSYYR79- dropped to about 20% after 10-min incubation with Lem3 and to less than 8% when incubation was extended to 30 min (Fig. 6B)." 1 ANKLE1 UNIPROT:Q8NAG6 activates GO:0000077 Phenotype 86a37944-c485-11e5-a92e-001a4ae51246 PMC4335191 When forced to the nucleus by transient inhibition of nuclear export, Ankle1 caused DNA cleavage and induced DNA damage response. 1 ANKLE1 UNIPROT:Q8NAG6 activates GO:0007059 Phenotype 996e68de-6afc-11e8-a34c-001a4a160176 PMC6007928 LEM-3 promotes proper chromosome segregation during meiotic cell division We next investigated the localization of LEM-3 using a strain expressing a GFP::LEM-3 fusion. 1 ANKLE1 UNIPROT:Q8NAG6 activates GO:0051168 Phenotype 58fff77a-2a6e-11e6-9884-001a4ae51246 PMC4888674 Scale bar: 5 μm Discussion discussion In this study we show that Ankle1 endonuclease shuttles between the cytoplasm and the nucleus by an active NLS-mediated nuclear import and a NES-mediated nuclear export, despite its predominant steady-state localization in the cytoplasm. 1 ANKLE1 UNIPROT:Q8NAG6 inhibits MESH:D010767 Phenotype 16daab26-bbdb-11e5-9b9d-001a4ae51247 PMC4322725 Indeed,in vitroassays demonstrated that Lem3 reverses the AnkX-dependent phosphocholination of Rab1 by removing the phosphocholine moiety from Rab1 (Tan et al., 2011;Table1). 1 ANKLE1 UNIPROT:Q8NAG6 inhibits GO:0043687 Phenotype 0d7a1532-c47b-11e5-91a7-001a4ae51247 PMC3248503 "The activity associated with total bacterial cell lysate in removing the PCylation signal from Rab1 suggested that Lem3 biochemically reverses the posttranslational modification on the target of AnkX." 1 ANKLE1 UNIPROT:Q8NAG6 inhibits GO:0051322 Phenotype ae9c4872-f0f4-11ea-93b9-001a4a160176 PMC7508374 We found that depletion of the conserved nuclease LEM-3/Ankle1 increases the frequency and persistence of anaphase bridging, consistent with its previously proposed role in processing erroneous recombination intermediates and correcting meiotic errors [34]. 1 ANKLE1 UNIPROT:Q8NAG6 activates GO:0006897 Phenotype 5d2cc6b0-bc4c-11e5-8abe-001a4ae51246 PMC2687517 Collectively, these findings indicate complex interactions among Pdr10, Pdr12, and Lem3, which produce secondary effects on Chs3 endocytosis. 1 ANKLE1 UNIPROT:Q8NAG6 activates MESH:D005690 Phenotype 0d7a1532-c47b-11e5-91a7-001a4ae51247 PMC3248503 Lem3 fully rescued the growth of the yeast strain harboringankXon galactose medium (Fig. 5B, strains iii and iv). 1 ANKLE1 UNIPROT:Q8NAG6 activates GO:0045332 Phenotype 182e41fa-c477-11e5-85e4-001a4ae51246 PMC4421010 Lem3 associates with the catalytic subunits encoded byDNF1andDNF2to produce the plasma membrane flippase. 1 UNIPROT:P48681 activates ANKLE1 UNIPROT:Q8NAG6 Protein 58fff77a-2a6e-11e6-9884-001a4ae51246 PMC4888674 Conclusions conclusion This study identifies a centrally locatedrev-type CRM1-dependent NES in the Ankle1 polypeptide and a C-terminal canonical mono-partite NLS, which together mediate shuttling of Ankle1 between the cytoplasm and the nucleus and maintain a predominantly cytoplasmic localization at steady state. 1 UNIPROT:Q9ZUT8 activates ANKLE1 UNIPROT:Q8NAG6 Protein 182e41fa-c477-11e5-85e4-001a4ae51246 PMC4421010 "To further probe the genetic relationship between the floppase activity produced by Pdr5 and Yor1 and the flippase activity dependent on Lem3, we constructed a series of strains lacking positive regulators of the flippase (Fig. 5B)." 1 CHEBI:16411 inhibits ANKLE1 UNIPROT:Q8NAG6 Chemical cbf41d20-c97a-11ee-b346-0050569a791b PMC10015115 Besides, both Lem1 and Lem3 proteins could be depleted by indole-3-acetic acid (IAA, auxin) in respective mutants, as shown by the disappearance of ∼130-kDa and ∼57-kDa bands in immunoblots, respectively (Fig. 3D). 1 UNIPROT:P39524 decreases ANKLE1 UNIPROT:Q8NAG6 Protein 799afdfe-c46b-11e5-85e4-001a4ae51246 25359886 We confirmed that the levels and localization of Drs2 and Dnf3 were comparable in these mutant cells and in wild-type cells (supplementary material Fig. S1B,C) and thus eliminated the possibility of increased levels of Lem3-independent flippases causing the reduction in the amount of exposed PtdEtn inopt2Δlem3Δ cells. 1 UNIPROT:Q12674 decreases ANKLE1 UNIPROT:Q8NAG6 Protein 799afdfe-c46b-11e5-85e4-001a4ae51246 25359886 We confirmed that the levels and localization of Drs2 and Dnf3 were comparable in these mutant cells and in wild-type cells (supplementary material Fig. S1B,C) and thus eliminated the possibility of increased levels of Lem3-independent flippases causing the reduction in the amount of exposed PtdEtn inopt2Δlem3Δ cells. 1 UNIPROT:Q12476 phosphorylatesProtein ANKLE1 UNIPROT:Q8NAG6 Protein 0ea5ae4c-ae96-11ec-89b1-0050569a791b PMCPMC8160109 However, it is unknown how LEM-3 phosphorylated by AIR-2 is recruited to the midbody. 1 UNIPROT:P53049 activates ANKLE1 UNIPROT:Q8NAG6 Protein 182e41fa-c477-11e5-85e4-001a4ae51246 PMC4421010 "To further probe the genetic relationship between the floppase activity produced by Pdr5 and Yor1 and the flippase activity dependent on Lem3, we constructed a series of strains lacking positive regulators of the flippase (Fig. 5B)." 1 UNIPROT:O14980 activates ANKLE1 UNIPROT:Q8NAG6 Protein 58fff77a-2a6e-11e6-9884-001a4ae51246 PMC4888674 Conclusions conclusion This study identifies a centrally locatedrev-type CRM1-dependent NES in the Ankle1 polypeptide and a C-terminal canonical mono-partite NLS, which together mediate shuttling of Ankle1 between the cytoplasm and the nucleus and maintain a predominantly cytoplasmic localization at steady state.