title: Defective nucleotide excision repair with normal centrosome structures and functions in the absence of all vertebrate centrins.
abstract: The principal microtubule-organizing center in animal cells, the centrosome, contains centrin, a small, conserved calcium-binding protein unique to eukaryotes. Several centrin isoforms exist and have been implicated in various cellular processes including nuclear export and deoxyribonucleic acid (DNA) repair. Although centrins are required for centriole/basal body duplication in lower eukaryotes, centrin functions in vertebrate centrosome duplication are less clear. To define these roles, we used gene targeting in the hyperrecombinogenic chicken DT40 cell line to delete all three centrin genes in individual clones. Unexpectedly, centrin-deficient cells underwent normal cellular division with no detectable cell cycle defects. Light and electron microscopy analyses revealed no significant difference in centrosome composition or ultrastructure. However, centrin deficiency made DT40 cells highly sensitive to ultraviolet (UV) irradiation, with Cetn3 deficiency exacerbating the sensitivity of Cetn4/Cetn2 double mutants. DNA damage checkpoints were intact, but repair of UV-induced DNA damage was delayed in centrin nulls. These data demonstrate a role for vertebrate centrin in nucleotide excision repair.
authors.affiliation: Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland Galway, Galway 091 524 411, Ireland.
authors.forename: Tiago J
authors.initials: TJ
authors.lastname: Dantas
authors.forename: Yifan
authors.initials: Y
authors.lastname: Wang
authors.forename: Pierce
authors.initials: P
authors.lastname: Lalor
authors.forename: Peter
authors.initials: P
authors.lastname: Dockery
authors.forename: Ciaran G
authors.initials: CG
authors.lastname: Morrison
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002455:Cell Division / Q000235:genetics / Q000528:radiation effects; D002460:Cell Line; D018385:Centrosome / Q000378:metabolism* / Q000528:radiation effects; D002645:Chickens; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics*; D055786:Gene Knockout Techniques; D014466:Ultraviolet Rays
chemical_list: D002135:Calcium-Binding Proteins; D002868:Chromosomal Proteins, Non-Histone; C056608:caltractin