H2A.Z depletion impairs proliferation and viability but not DNA double-strand breaks repair in human immortalized and tumoral cell lines.
In mammalian cells, DNA double-strand breaks (DSB) can be repaired by 2 main pathways, homologous recombination (HR) and non-homologous end joining (NHEJ). To give access to DNA damage to the repair machinery the chromatin structure needs to be relaxed, and chromatin modifications play major roles in the control of these processes. Among the chromatin modifications, changes in nucleosome composition can influence DNA damage response as observed with the H2A.Z histone variant in yeast. In mammals, p400, an ATPase of the SWI/SNF family able to incorporate H2A.Z in chromatin, was found to be important for histone ubiquitination and BRCA1 recruitment around DSB or for HR in cooperation with Rad51. Recent data with 293T cells showed that mammalian H2A.Z is recruited to DSBs and is important to control DNA resection, therefore participating both in HR and NHEJ. Here we show that depletion of H2A.Z in the osteosarcoma U2OS cell line and in immortalized human fibroblasts does not change parameters of DNA DSB repair while affecting clonogenic ability and cell cycle distribution. In addition, no recruitment of H2A.Z around DSB can be detected in U2OS cells either after local laser irradiation or by chromatin immunoprecipitation. These data suggest that the role of H2A.Z in DSB repair is not ubiquitous in mammals. In addition, given that important cellular parameters, such as cell viability and cell cycle distribution, are more sensitive to H2A.Z depletion than DNA repair, our results underline the difficulty to investigate the role of versatile factors such as H2A.Z.
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Gemael-Cedrick
GC
Taty-Taty
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Celine
C
Courilleau
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Muriel
M
Quaranta
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Alexandre
A
Carayon
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Catherine
C
Chailleux
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
François
F
Aymard
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Didier
D
Trouche
Université de Toulouse; UPS; LBCMCP; Toulouse, France; CNRS; LBCMCP; Toulouse, France.
Yvan
Y
Canitrot
D059447:Cell Cycle Checkpoints / Q000235:genetics; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D006657:Histones / Q000235:genetics*; D006801:Humans; D007834:Lasers
D006657:Histones; C077020:histone H2A.F-Z
DNA repair; H2A.Z; NHEJ; chromatin; homologous recombination; p400