Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis

Jingjing Jiang; Lu Bai; Jennifer A. Surtees; Zekeriyya Gemici; Michelle D. Wang; Eric Alani

doi:10.1016/j.molcel.2005.10.014

Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis

Jingjing Jiang, Lu Bai, Jennifer A. Surtees, Zekeriyya Gemici, Michelle D. Wang, Eric Alani

Biochemistry & Molecular Biology

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

50 Scopus citations

Abstract

Mismatch repair (MMR) is initiated by MutS family proteins (MSH) that recognize DNA mismatches and recruit downstream repair factors. We used a single-molecule DNA-unzipping assay to probe interactions between S. cerevisiae MSH2-MSH6 and a variety of DNA mismatch substrates. This work revealed a high-specificity binding state of MSH proteins for mismatch DNA that was not observed in bulk assays and allowed us to measure the affinity of MSH2-MSH6 for mismatch DNA as well as its footprint on DNA surrounding the mismatch site. Unzipping analysis with mismatch substrates containing an end blocked by lac repressor allowed us to identify MSH proteins present on DNA between the mismatch and the block, presumably in an ATP-dependent sliding clamp mode. These studies provide a high-resolution approach to study MSH interactions with DNA mismatches and supply evidence to support and refute different models proposed for initiation steps in MMR.

Original language	English (US)
Pages (from-to)	771-781
Number of pages	11
Journal	Molecular cell
Volume	20
Issue number	5
DOIs	https://doi.org/10.1016/j.molcel.2005.10.014
State	Published - Dec 9 2005

All Science Journal Classification (ASJC) codes

Molecular Biology
Cell Biology

Access to Document

10.1016/j.molcel.2005.10.014

Cite this

@article{81bf4b4b47d340bab7b8e1316cb6ef26,

title = "Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis",

abstract = "Mismatch repair (MMR) is initiated by MutS family proteins (MSH) that recognize DNA mismatches and recruit downstream repair factors. We used a single-molecule DNA-unzipping assay to probe interactions between S. cerevisiae MSH2-MSH6 and a variety of DNA mismatch substrates. This work revealed a high-specificity binding state of MSH proteins for mismatch DNA that was not observed in bulk assays and allowed us to measure the affinity of MSH2-MSH6 for mismatch DNA as well as its footprint on DNA surrounding the mismatch site. Unzipping analysis with mismatch substrates containing an end blocked by lac repressor allowed us to identify MSH proteins present on DNA between the mismatch and the block, presumably in an ATP-dependent sliding clamp mode. These studies provide a high-resolution approach to study MSH interactions with DNA mismatches and supply evidence to support and refute different models proposed for initiation steps in MMR.",

author = "Jingjing Jiang and Lu Bai and Surtees, {Jennifer A.} and Zekeriyya Gemici and Wang, {Michelle D.} and Eric Alani",

note = "Funding Information: We thank Alla Shundrovsky, Jianyong Tang, and Micheal Mwangi for technical advice; Tatiana Velikodvorskaya and Robert Weissberg for providing lac repressor; and Eric Greene, John Lis, and Jeff Roberts for helpful comments and technical advice. J.J. and E.A. were supported by NIH research grant GM53085 and Z.G. by a Cornell Presidential Research Scholars Fellowship. J.A.S. is a Research Fellow of the National Cancer Institute of Canada supported with funds provided by the Terry Fox Run. M.D.W. was supported by NIH research grant GM59849 and the Keck Foundation's Distinguished Young Scholar Award. ",

year = "2005",

month = dec,

day = "9",

doi = "10.1016/j.molcel.2005.10.014",

language = "English (US)",

volume = "20",

pages = "771--781",

journal = "Molecular cell",

issn = "1097-2765",

publisher = "Cell Press",

number = "5",

}

TY - JOUR

T1 - Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis

AU - Jiang, Jingjing

AU - Bai, Lu

AU - Surtees, Jennifer A.

AU - Gemici, Zekeriyya

AU - Wang, Michelle D.

AU - Alani, Eric

N1 - Funding Information: We thank Alla Shundrovsky, Jianyong Tang, and Micheal Mwangi for technical advice; Tatiana Velikodvorskaya and Robert Weissberg for providing lac repressor; and Eric Greene, John Lis, and Jeff Roberts for helpful comments and technical advice. J.J. and E.A. were supported by NIH research grant GM53085 and Z.G. by a Cornell Presidential Research Scholars Fellowship. J.A.S. is a Research Fellow of the National Cancer Institute of Canada supported with funds provided by the Terry Fox Run. M.D.W. was supported by NIH research grant GM59849 and the Keck Foundation's Distinguished Young Scholar Award.

PY - 2005/12/9

Y1 - 2005/12/9

N2 - Mismatch repair (MMR) is initiated by MutS family proteins (MSH) that recognize DNA mismatches and recruit downstream repair factors. We used a single-molecule DNA-unzipping assay to probe interactions between S. cerevisiae MSH2-MSH6 and a variety of DNA mismatch substrates. This work revealed a high-specificity binding state of MSH proteins for mismatch DNA that was not observed in bulk assays and allowed us to measure the affinity of MSH2-MSH6 for mismatch DNA as well as its footprint on DNA surrounding the mismatch site. Unzipping analysis with mismatch substrates containing an end blocked by lac repressor allowed us to identify MSH proteins present on DNA between the mismatch and the block, presumably in an ATP-dependent sliding clamp mode. These studies provide a high-resolution approach to study MSH interactions with DNA mismatches and supply evidence to support and refute different models proposed for initiation steps in MMR.

AB - Mismatch repair (MMR) is initiated by MutS family proteins (MSH) that recognize DNA mismatches and recruit downstream repair factors. We used a single-molecule DNA-unzipping assay to probe interactions between S. cerevisiae MSH2-MSH6 and a variety of DNA mismatch substrates. This work revealed a high-specificity binding state of MSH proteins for mismatch DNA that was not observed in bulk assays and allowed us to measure the affinity of MSH2-MSH6 for mismatch DNA as well as its footprint on DNA surrounding the mismatch site. Unzipping analysis with mismatch substrates containing an end blocked by lac repressor allowed us to identify MSH proteins present on DNA between the mismatch and the block, presumably in an ATP-dependent sliding clamp mode. These studies provide a high-resolution approach to study MSH interactions with DNA mismatches and supply evidence to support and refute different models proposed for initiation steps in MMR.

UR - http://www.scopus.com/inward/record.url?scp=28444499354&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28444499354&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2005.10.014

DO - 10.1016/j.molcel.2005.10.014

M3 - Article

C2 - 16337600

AN - SCOPUS:28444499354

SN - 1097-2765

VL - 20

SP - 771

EP - 781

JO - Molecular cell

JF - Molecular cell

IS - 5

ER -

Detection of high-affinity and sliding clamp modes for MSH2-MSH6 by single-molecule unzipping force analysis

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this