Eukaryotic DNA mismatch repair in vitro

Fenghua Yuan; Limin Song; Fengsong Liu; Liya Gu; Yanbin Zhang

doi:10.1007/978-1-61779-998-3_11

Eukaryotic DNA mismatch repair in vitro

Fenghua Yuan, Limin Song, Fengsong Liu, Liya Gu, Yanbin Zhang

Research output: Chapter in Book/Report/Conference proceeding › Chapter

3 Scopus citations

Abstract

Mismatch repair corrects biosynthetic errors generated during DNA replication. Mismatch repair deficiency causes a mutator phenotype and directly underlies hereditary nonpolyposis colorectal cancer and some sporadic cancers. Because of remarkably high conservation of the mismatch repair machinery between the budding yeast (Saccharomyces cerevisiae) and humans, the study of mismatch repair in yeast has provided tremendous insights into the mechanisms of this repair pathway in humans. Here we describe a set of practical protocols for how to prepare the yeast and HeLa cell-free nuclear extracts and site-specific DNA mismatch substrates, and how to carry out the in vitro mismatch repair assay. We validated the yeast cell-free system by the mismatch repair deficient strain (Δmsh2) and the complementation assay with purified yeast MutSα.

Original language	English (US)
Title of host publication	DNA Repair Protocols
Publisher	Humana Press Inc.
Pages	149-162
Number of pages	14
ISBN (Print)	9781617799976
DOIs	https://doi.org/10.1007/978-1-61779-998-3_11
State	Published - 2012

Publication series

Name	Methods in Molecular Biology
Volume	920
ISSN (Print)	1064-3745

Keywords

Cancer
Cell-free nuclear extract
Human
In vitro DNA mismatch repair
MMR
Saccharomyces cerevisiae
Yeast

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-61779-998-3_11

Cite this

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title = "Eukaryotic DNA mismatch repair in vitro",

abstract = "Mismatch repair corrects biosynthetic errors generated during DNA replication. Mismatch repair deficiency causes a mutator phenotype and directly underlies hereditary nonpolyposis colorectal cancer and some sporadic cancers. Because of remarkably high conservation of the mismatch repair machinery between the budding yeast (Saccharomyces cerevisiae) and humans, the study of mismatch repair in yeast has provided tremendous insights into the mechanisms of this repair pathway in humans. Here we describe a set of practical protocols for how to prepare the yeast and HeLa cell-free nuclear extracts and site-specific DNA mismatch substrates, and how to carry out the in vitro mismatch repair assay. We validated the yeast cell-free system by the mismatch repair deficient strain (Δmsh2) and the complementation assay with purified yeast MutSα.",

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T1 - Eukaryotic DNA mismatch repair in vitro

AU - Yuan, Fenghua

AU - Song, Limin

AU - Liu, Fengsong

AU - Gu, Liya

AU - Zhang, Yanbin

PY - 2012

Y1 - 2012

N2 - Mismatch repair corrects biosynthetic errors generated during DNA replication. Mismatch repair deficiency causes a mutator phenotype and directly underlies hereditary nonpolyposis colorectal cancer and some sporadic cancers. Because of remarkably high conservation of the mismatch repair machinery between the budding yeast (Saccharomyces cerevisiae) and humans, the study of mismatch repair in yeast has provided tremendous insights into the mechanisms of this repair pathway in humans. Here we describe a set of practical protocols for how to prepare the yeast and HeLa cell-free nuclear extracts and site-specific DNA mismatch substrates, and how to carry out the in vitro mismatch repair assay. We validated the yeast cell-free system by the mismatch repair deficient strain (Δmsh2) and the complementation assay with purified yeast MutSα.

AB - Mismatch repair corrects biosynthetic errors generated during DNA replication. Mismatch repair deficiency causes a mutator phenotype and directly underlies hereditary nonpolyposis colorectal cancer and some sporadic cancers. Because of remarkably high conservation of the mismatch repair machinery between the budding yeast (Saccharomyces cerevisiae) and humans, the study of mismatch repair in yeast has provided tremendous insights into the mechanisms of this repair pathway in humans. Here we describe a set of practical protocols for how to prepare the yeast and HeLa cell-free nuclear extracts and site-specific DNA mismatch substrates, and how to carry out the in vitro mismatch repair assay. We validated the yeast cell-free system by the mismatch repair deficient strain (Δmsh2) and the complementation assay with purified yeast MutSα.

KW - Cancer

KW - Cell-free nuclear extract

KW - Human

KW - In vitro DNA mismatch repair

KW - MMR

KW - Saccharomyces cerevisiae

KW - Yeast

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ER -

Eukaryotic DNA mismatch repair in vitro

Abstract

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Keywords

ASJC Scopus subject areas

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