Ku is a 5′-dRP/AP lyase that excises nucleotide damage near broken ends

Steven A. Roberts, Natasha Strande, Martin D. Burkhalter, Christina Strom, Jody M. Havener, Paul Hasty, Dale A. Ramsden

Research output: Contribution to journalArticlepeer-review

126 Scopus citations

Abstract

Mammalian cells require non-homologous end joining (NHEJ) for the efficient repair of chromosomal DNA double-strand breaks. A key feature of biological sources of strand breaks is associated nucleotide damage, including base loss (abasic or apurinic/apyrimidinic (AP) sites). At single-strand breaks, 5′-terminal abasic sites are excised by the 5′-deoxyribose-5- phosphate (5′-dRP) lyase activity of DNA polymerase Β (pol Β): here we show, in vitro and in cells, that accurate and efficient repair by NHEJ of double-strand breaks with such damage similarly requires 5′-dRP/AP lyase activity. Classically defined NHEJ is moreover uniquely effective at coupling this end-cleaning step to joining in cells, helping to distinguish this pathway from otherwise robust alternative NHEJ pathways. The NHEJ factor Ku can be identified as an effective 5′-dRP/AP lyase. In a similar manner to other lyases, Ku nicks DNA 3′ of an abasic site by a mechanism involving a Schiff-base covalent intermediate with the abasic site. We show by using cell extracts that Ku is essential for the efficient removal of AP sites near double-strand breaks and, consistent with this result, that joining of such breaks is specifically decreased in cells complemented with a lyase-attenuated Ku mutant. Ku had previously been presumed only to recognize ends and recruit other factors that process ends; our data support an unexpected direct role for Ku in end-processing steps as well.

Original languageEnglish (US)
Pages (from-to)1214-1217
Number of pages4
JournalNature
Volume464
Issue number7292
DOIs
StatePublished - Apr 22 2010

ASJC Scopus subject areas

  • General

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