C-NAP1 and rootletin restrain DNA damage-induced centriole splitting and facilitate ciliogenesis.
Cilia are found on most human cells and exist as motile cilia or non-motile primary cilia. Primary cilia play sensory roles in transducing various extracellular signals, and defective ciliary functions are involved in a wide range of human diseases. Centrosomes are the principal microtubule-organizing centers of animal cells and contain two centrioles. We observed that DNA damage causes centriole splitting in non-transformed human cells, with isolated centrioles carrying the mother centriole markers CEP170 and ninein but not kizuna or cenexin. Loss of centriole cohesion through siRNA depletion of C-NAP1 or rootletin increased radiation-induced centriole splitting, with C-NAP1-depleted isolated centrioles losing mother markers. As the mother centriole forms the basal body in primary cilia, we tested whether centriole splitting affected ciliogenesis. While irradiated cells formed apparently normal primary cilia, most cilia arose from centriolar clusters, not from isolated centrioles. Furthermore, C-NAP1 or rootletin knockdown reduced primary cilium formation. Therefore, the centriole cohesion apparatus at the proximal end of centrioles may provide a target that can affect primary cilium formation as part of the DNA damage response.
Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.
Pauline C
PC
Conroy
Chiara
C
Saladino
Tiago J
TJ
Dantas
Pierce
P
Lalor
Peter
P
Dockery
Ciaran G
CG
Morrison
D015415:Biomarkers / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002502:Centrioles / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects; D002868:Chromosomal Proteins, Non-Histone; D002923:Cilia / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects; D003598:Cytoskeletal Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage; D005720:Gamma Rays; D020868:Gene Silencing; D006801:Humans; D008869:Microtubule-Associated Proteins; D009024:Morphogenesis / Q000502:physiology*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000528:radiation effects
D015415:Biomarkers; C499237:CROCC protein, human; D018797:Cell Cycle Proteins; C502822:Cep170 protein, human; D002868:Chromosomal Proteins, Non-Histone; D003598:Cytoskeletal Proteins; D008869:Microtubule-Associated Proteins; C413383:NCAPD2 protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D000075223:Poly-ADP-Ribose Binding Proteins; D034741:RNA, Small Interfering