CAF-1 is essential for Drosophila development and involved in the maintenance of epigenetic memory.
DNA synthesis during S-phase and upon DNA repair is accompanied by chromatin assembly. The chromatin assembly factor CAF-1 has been biochemically well-characterized to deposit histones onto newly synthesized DNA. To gain insights into the in vivo functions of CAF-1 in Drosophila, we generated null mutants of the largest subunit of dCAF-1, dCAF-1-p180. We show that, unlike CAF-1 mutant yeast, dCAF-1-p180 mutant flies are hemizygous lethal. Removal of maternal dCAF-1-p180 activity by germline clones blocks oogenesis. Tissue-specific deletion of dCAF-1-p180 in the eye primordia disrupts eye development. In addition, reduction of dCAF-1-p180 activity suppresses gene silencing at heterochromatin and antagonizes Polycomb-mediated cell fate determination. Furthermore, heterozygous dCAF-1-p180 mutant flies display an increased sensitivity to gamma-irradiation and a reduced efficiency in recombinational double strand break (DSB) repair. Our experiments also show that human hCAF-1-p150 can rescue the dCAF-1-p180 mutant flies, demonstrating a functional conservation of eukaryotic CAF-1 activities in vivo. Together, our results establish that dCAF-1-p180 is an essential gene for Drosophila development and further underscore the importance of dCAF-1 in regulating gene expression and DNA repair in vivo.
Song
Yanjun
Y
National Laboratory of Biomacromolecules and State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, The Chinese Academy of Sciences, Datun Road 15, Chaoyang District, Beijing 100101, China.
He
Feng
F
Xie
Gengqiang
G
Guo
Xiaoyan
X
Xu
Yanjuan
Y
Chen
Yixu
Y
Liang
Xuehong
X
Stagljar
Igor
I
Egli
Dieter
D
Ma
Jun
J
Jiao
Renjie
R
D000818:Animals; D030801:Animals, Genetically Modified; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000196:embryology* / Q000378:metabolism; D044127:Epigenesis, Genetic; D005123:Eye / Q000196:embryology; D005260:Female; D020868:Gene Silencing; D008297:Male; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis; D056505:Retinoblastoma-Binding Protein 4; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
C489780:CNOT8 protein, human; C110251:Caf1-55 protein, Drosophila; D002868:Chromosomal Proteins, Non-Histone; D029721:Drosophila Proteins; D018832:Molecular Chaperones; D056505:Retinoblastoma-Binding Protein 4; D014157:Transcription Factors