Abstract

We are currently engaged in study of several aspects of site specific cleavage and methylation of DNA. The E. coli RI DNA restriction and modification enzymes, are being analyzed as a model system for study of sequence-specific DNA protein interaction. The interaction of the EcoRI enzymes with short DNA molecules will be investigated with the aim of obtaining thermodynamic and kinetic parameters governing the interaction. Utilizing absorbance and flourescence methods, we hope to apply rapid kinetic methods to analysis of such interactions. We have cloned the structural genes for the EcoRI enzymes and have determined their DNA sequence. Hence, we have obtained primary sequences of the two proteins. By employing site specific mutagenesis and DNA sequencing techniques we hope to establish a structure-function map for the two proteins. We are also pursuing work of the mechanism and biological function of the Escherichia coli dam methylase which is responsible for almost all 6-methyladenine residue in this organism.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM023719-10
Application #
3271854
Study Section
Biochemistry Study Section (BIO)
Project Start
1977-08-01
Project End
1986-07-31
Budget Start
1985-08-01
Budget End
1986-07-31
Support Year
10
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705

  Publications

Modrich, Paul (2016) Mechanisms in E. coli and Human Mismatch Repair (Nobel Lecture). Angew Chem Int Ed Engl 55:8490-501
Pluciennik, Anna; Burdett, Vickers; Lukianova, Olga et al. (2009) Involvement of the beta clamp in methyl-directed mismatch repair in vitro. J Biol Chem 284:32782-91
Pluciennik, Anna; Modrich, Paul (2007) Protein roadblocks and helix discontinuities are barriers to the initiation of mismatch repair. Proc Natl Acad Sci U S A 104:12709-13
Lopez de Saro, Francisco J; Marinus, Martin G; Modrich, Paul et al. (2006) The beta sliding clamp binds to multiple sites within MutL and MutS. J Biol Chem 281:14340-9
Iyer, Ravi R; Pluciennik, Anna; Burdett, Vickers et al. (2006) DNA mismatch repair: functions and mechanisms. Chem Rev 106:302-23
Bjornson, Keith P; Blackwell, Leonard J; Sage, Harvey et al. (2003) Assembly and molecular activities of the MutS tetramer. J Biol Chem 278:34667-73
Bjornson, Keith P; Modrich, Paul (2003) Differential and simultaneous adenosine di- and triphosphate binding by MutS. J Biol Chem 278:18557-62
Baitinger, Celia; Burdett, Vickers; Modrich, Paul (2003) Hydrolytically deficient MutS E694A is defective in the MutL-dependent activation of MutH and in the mismatch-dependent assembly of the MutS.MutL.heteroduplex complex. J Biol Chem 278:49505-11
Viswanathan, M; Burdett, V; Baitinger, C et al. (2001) Redundant exonuclease involvement in Escherichia coli methyl-directed mismatch repair. J Biol Chem 276:31053-8
Blackwell, L J; Bjornson, K P; Allen, D J et al. (2001) Distinct MutS DNA-binding modes that are differentially modulated by ATP binding and hydrolysis. J Biol Chem 276:34339-47

Showing the most recent 10 out of 46 publications