title: Nucleo-cytoplasmic shuttling of the endonuclease ankyrin repeats and LEM domain-containing protein 1 (Ankle1) is mediated by canonical nuclear export- and nuclear import signals.
abstract: Ankyrin repeats and LEM domain containing protein 1 (Ankle1) belongs to the LEM protein family, whose members share a chromatin-interacting LEM motif. Unlike most other LEM proteins, Ankle1 is not an integral protein of the inner nuclear membrane but shuttles between the nucleus and the cytoplasm. It contains a GIY-YIG-type nuclease domain, but its function is unknown. The mammalian genome encodes only one other GIY-YIG domain protein, termed Slx1. Slx1 has been described as a resolvase that processes Holliday junctions during homologous recombination-mediated DNA double strand break repair. Resolvase activity is regulated in a spatial and temporal manner during the cell cycle. We hypothesized that Ankle1 may have a similar function and its nucleo-cytoplasmic shuttling may contribute to the regulation of Ankle1 activity. Hence, we aimed at identifying the domains mediating Ankle1 shuttling and investigating whether cellular localization is affected during DNA damage response.
  
  
  
  Sequence analysis predicts the presence of two canonical nuclear import and export signals in Ankle1. Immunofluorescence microscopy of cells expressing wild-type and various mutated Ankle1-fusion proteins revealed a C-terminally located classical monopartite nuclear localization signal and a centrally located CRM1-dependent nuclear export signal that mediate nucleo-cytoplasmic shuttling of Ankle1. These sequences are also functional in heterologous proteins. The predominant localization of Ankle1 in the cytoplasm, however, does not change upon induction of several DNA damage response pathways throughout the cell cycle.
  
  
  
  We identified the domains mediating nuclear import and export of Ankle1. Ankle1's cellular localization was not affected following DNA damage.
authors.affiliation: Max F. Perutz Laboratories (MFPL), Department of Medical Biochemistry, Medical University of Vienna, Vienna Biocenter (VBC), Vienna, Austria.
authors.forename: Livija
authors.initials: L
authors.lastname: Zlopasa
authors.affiliation: Max F. Perutz Laboratories (MFPL), Department of Medical Biochemistry, Medical University of Vienna, Vienna Biocenter (VBC), Vienna, Austria. andreas.brachner@univie.ac.at.
authors.forename: Andreas
authors.initials: A
authors.lastname: Brachner
authors.affiliation: Max F. Perutz Laboratories (MFPL), Department of Medical Biochemistry, Medical University of Vienna, Vienna Biocenter (VBC), Vienna, Austria. roland.foisner@meduniwien.ac.at.
authors.forename: Roland
authors.initials: R
authors.lastname: Foisner
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D004249:DNA Damage; D004252:DNA Mutational Analysis; D004720:Endonucleases / Q000737:chemistry / Q000378:metabolism*; D017930:Genes, Reporter; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals / Q000378:metabolism*; D000072417:Protein Domains
chemical_list: D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; C579363:ANKLE1 protein, human; D004720:Endonucleases; D006918:Hydroxyurea
keywords: Ankle1; LEM protein; Nuclear export; Nuclear transport; Nuclease; Resolvase