TY - JOUR
T1 - Pre-existing H4K16ac levels in euchromatin drive DNA repair by homologous recombination in S-phase
AU - Horikoshi, Nobuo
AU - Sharma, Dharmendra
AU - Leonard, Fransisca
AU - Pandita, Raj K.
AU - Charaka, Vijaya K.
AU - Hambarde, Shashank
AU - Horikoshi, Nobuko T.
AU - Khaitan, Puja G.
AU - Chakraborty, Sharmistha
AU - Cote, Jacques
AU - Godin, Biana
AU - Hunt, Clayton R.
AU - Pandita, Tej K.
N1 - Funding Information:
We thank members of Pandita laboratory for discussions during the inception and the completion of this study, respectively. Thanks are due to Drs Albino Bacolla and Mauro Ferrari for their help. This study is supported the Houston Methodist Research Institute and NIH RO1 CA129537 and RO1 GM109768 to TKP.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - The homologous recombination (HR) repair pathway maintains genetic integrity after DNA double-strand break (DSB) damage and is particularly crucial for maintaining fidelity of expressed genes. Histone H4 acetylation on lysine 16 (H4K16ac) is associated with transcription, but how pre-existing H4K16ac directly affects DSB repair is not known. To answer this question, we used CRISPR/Cas9 technology to introduce I-SceI sites, or repair pathway reporter cassettes, at defined locations within gene-rich (high H4K16ac/euchromatin) and gene-poor (low H4K16ac/heterochromatin) regions. The frequency of DSB repair by HR is higher in gene-rich regions. Interestingly, artificially targeting H4K16ac at specific locations using gRNA/dCas9-MOF increases HR frequency in euchromatin. Finally, inhibition/depletion of RNA polymerase II or Cockayne syndrome B protein leads to decreased recruitment of HR factors at DSBs. These results indicate that the pre-existing H4K16ac status at specific locations directly influences the repair of local DNA breaks, favoring HR in part through the transcription machinery.
AB - The homologous recombination (HR) repair pathway maintains genetic integrity after DNA double-strand break (DSB) damage and is particularly crucial for maintaining fidelity of expressed genes. Histone H4 acetylation on lysine 16 (H4K16ac) is associated with transcription, but how pre-existing H4K16ac directly affects DSB repair is not known. To answer this question, we used CRISPR/Cas9 technology to introduce I-SceI sites, or repair pathway reporter cassettes, at defined locations within gene-rich (high H4K16ac/euchromatin) and gene-poor (low H4K16ac/heterochromatin) regions. The frequency of DSB repair by HR is higher in gene-rich regions. Interestingly, artificially targeting H4K16ac at specific locations using gRNA/dCas9-MOF increases HR frequency in euchromatin. Finally, inhibition/depletion of RNA polymerase II or Cockayne syndrome B protein leads to decreased recruitment of HR factors at DSBs. These results indicate that the pre-existing H4K16ac status at specific locations directly influences the repair of local DNA breaks, favoring HR in part through the transcription machinery.
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U2 - 10.1038/s42003-019-0498-z
DO - 10.1038/s42003-019-0498-z
M3 - Article
C2 - 31925071
AN - SCOPUS:85071153719
SN - 2399-3642
VL - 2
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 253
ER -