Mre11gRad50gXrs2 and Sae2 promote 5′ strand resection of DNA double-strand breaks

Matthew L. Nicolette; Kihoon Lee; Zhi Guo; Mridula Rani; Julia M. Chow; Sang Eun Lee; Tanya T. Paull

doi:10.1038/nsmb.1957

Mre11gRad50gXrs2 and Sae2 promote 5′ strand resection of DNA double-strand breaks

Matthew L. Nicolette, Kihoon Lee, Zhi Guo, Mridula Rani, Julia M. Chow, Sang Eun Lee, Tanya T. Paull

Molecular Medicine

Research output: Contribution to journal › Article › peer-review

151 Scopus citations

Abstract

The repair of DNA double-strand breaks (DSBs) by homologous recombination is essential for genomic stability. The first step in this process is resection of 5′ strands to generate 3′ single-stranded DNA intermediates. Efficient resection in budding yeast requires the Mre11gRad50gXrs2 (MRX) complex and the Sae2 protein, although the role of MRX has been unclear because Mre11 paradoxically has 3′ → 5′ exonuclease activity in vitro. Here we reconstitute resection with purified MRX, Sae2 and Exo1 proteins and show that degradation of the 5′ strand is catalyzed by Exo1 yet completely dependent on MRX and Sae2 when Exo1 levels are limiting. This stimulation is mainly caused by cooperative binding of DNA substrates by Exo1, MRX and Sae2. This work establishes the direct role of MRX and Sae2 in promoting the resection of 5′ strands in DNA DSB repair.

Original language	English (US)
Pages (from-to)	1478-1485
Number of pages	8
Journal	Nature Structural and Molecular Biology
Volume	17
Issue number	12
DOIs	https://doi.org/10.1038/nsmb.1957
State	Published - Dec 2010

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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title = "Mre11gRad50gXrs2 and Sae2 promote 5′ strand resection of DNA double-strand breaks",

abstract = "The repair of DNA double-strand breaks (DSBs) by homologous recombination is essential for genomic stability. The first step in this process is resection of 5′ strands to generate 3′ single-stranded DNA intermediates. Efficient resection in budding yeast requires the Mre11gRad50gXrs2 (MRX) complex and the Sae2 protein, although the role of MRX has been unclear because Mre11 paradoxically has 3′ → 5′ exonuclease activity in vitro. Here we reconstitute resection with purified MRX, Sae2 and Exo1 proteins and show that degradation of the 5′ strand is catalyzed by Exo1 yet completely dependent on MRX and Sae2 when Exo1 levels are limiting. This stimulation is mainly caused by cooperative binding of DNA substrates by Exo1, MRX and Sae2. This work establishes the direct role of MRX and Sae2 in promoting the resection of 5′ strands in DNA DSB repair.",

author = "Nicolette, {Matthew L.} and Kihoon Lee and Zhi Guo and Mridula Rani and Chow, {Julia M.} and Lee, {Sang Eun} and Paull, {Tanya T.}",

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AU - Nicolette, Matthew L.

AU - Lee, Kihoon

AU - Guo, Zhi

AU - Rani, Mridula

AU - Chow, Julia M.

AU - Lee, Sang Eun

AU - Paull, Tanya T.

PY - 2010/12

Y1 - 2010/12

N2 - The repair of DNA double-strand breaks (DSBs) by homologous recombination is essential for genomic stability. The first step in this process is resection of 5′ strands to generate 3′ single-stranded DNA intermediates. Efficient resection in budding yeast requires the Mre11gRad50gXrs2 (MRX) complex and the Sae2 protein, although the role of MRX has been unclear because Mre11 paradoxically has 3′ → 5′ exonuclease activity in vitro. Here we reconstitute resection with purified MRX, Sae2 and Exo1 proteins and show that degradation of the 5′ strand is catalyzed by Exo1 yet completely dependent on MRX and Sae2 when Exo1 levels are limiting. This stimulation is mainly caused by cooperative binding of DNA substrates by Exo1, MRX and Sae2. This work establishes the direct role of MRX and Sae2 in promoting the resection of 5′ strands in DNA DSB repair.

AB - The repair of DNA double-strand breaks (DSBs) by homologous recombination is essential for genomic stability. The first step in this process is resection of 5′ strands to generate 3′ single-stranded DNA intermediates. Efficient resection in budding yeast requires the Mre11gRad50gXrs2 (MRX) complex and the Sae2 protein, although the role of MRX has been unclear because Mre11 paradoxically has 3′ → 5′ exonuclease activity in vitro. Here we reconstitute resection with purified MRX, Sae2 and Exo1 proteins and show that degradation of the 5′ strand is catalyzed by Exo1 yet completely dependent on MRX and Sae2 when Exo1 levels are limiting. This stimulation is mainly caused by cooperative binding of DNA substrates by Exo1, MRX and Sae2. This work establishes the direct role of MRX and Sae2 in promoting the resection of 5′ strands in DNA DSB repair.

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