Classical non-homologous end-joining pathway utilizes nascent RNA for error-free double-strand break repair of transcribed genes

Anirban Chakraborty, Nisha Tapryal, Tatiana Venkova, Nobuo Horikoshi, Raj K. Pandita, Altaf H. Sarker, Partha Sarkar, Tej K. Pandita, Tapas Hazra

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

DNA double-strand breaks (DSBs) leading to loss of nucleotides in the transcribed region can be lethal. Classical non-homologous end-joining (C-NHEJ) is the dominant pathway for DSB repair (DSBR) in adult mammalian cells. Here we report that during such DSBR, mammalian C-NHEJ proteins form a multiprotein complex with RNA polymerase II and preferentially associate with the transcribed genes after DSB induction. Depletion of C-NHEJ factors significantly abrogates DSBR in transcribed but not in non-transcribed genes. We hypothesized that nascent RNA can serve as a template for restoring the missing sequences, thus allowing error-free DSBR. We indeed found pre-mRNA in the C-NHEJ complex. Finally, when a DSB-containing plasmid with several nucleotides deleted within the E. coli lacZ gene was allowed time to repair in lacZ-expressing mammalian cells, a functional lacZ plasmid could be recovered from control but not C-NHEJ factor-depleted cells, providing important mechanistic insights into C-NHEJ-mediated error-free DSBR of the transcribed genome.

Original languageEnglish (US)
Article number13049
JournalNature Communications
Volume7
DOIs
StatePublished - Oct 5 2016

Fingerprint

Joining
genes
strands
Repair
Genes
RNA
Plasmids
plasmids
Nucleotides
nucleotides
Multiprotein Complexes
Lac Operon
Double-Stranded DNA Breaks
RNA Polymerase II
RNA Precursors
Cells
genome
Genome
Escherichia coli
1,2-di-(4-sulfamidophenyl)-4-butylpyrazolidine-3,5-dione

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Classical non-homologous end-joining pathway utilizes nascent RNA for error-free double-strand break repair of transcribed genes. / Chakraborty, Anirban; Tapryal, Nisha; Venkova, Tatiana; Horikoshi, Nobuo; Pandita, Raj K.; Sarker, Altaf H.; Sarkar, Partha; Pandita, Tej K.; Hazra, Tapas.

In: Nature Communications, Vol. 7, 13049, 05.10.2016.

Research output: Contribution to journalArticle

Chakraborty, Anirban ; Tapryal, Nisha ; Venkova, Tatiana ; Horikoshi, Nobuo ; Pandita, Raj K. ; Sarker, Altaf H. ; Sarkar, Partha ; Pandita, Tej K. ; Hazra, Tapas. / Classical non-homologous end-joining pathway utilizes nascent RNA for error-free double-strand break repair of transcribed genes. In: Nature Communications. 2016 ; Vol. 7.
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abstract = "DNA double-strand breaks (DSBs) leading to loss of nucleotides in the transcribed region can be lethal. Classical non-homologous end-joining (C-NHEJ) is the dominant pathway for DSB repair (DSBR) in adult mammalian cells. Here we report that during such DSBR, mammalian C-NHEJ proteins form a multiprotein complex with RNA polymerase II and preferentially associate with the transcribed genes after DSB induction. Depletion of C-NHEJ factors significantly abrogates DSBR in transcribed but not in non-transcribed genes. We hypothesized that nascent RNA can serve as a template for restoring the missing sequences, thus allowing error-free DSBR. We indeed found pre-mRNA in the C-NHEJ complex. Finally, when a DSB-containing plasmid with several nucleotides deleted within the E. coli lacZ gene was allowed time to repair in lacZ-expressing mammalian cells, a functional lacZ plasmid could be recovered from control but not C-NHEJ factor-depleted cells, providing important mechanistic insights into C-NHEJ-mediated error-free DSBR of the transcribed genome.",
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