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Emeritus Professor of Biochemistry and Biophysics
Ph.D. University of Birmingham, UK 1959
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Molecular Biology of DNA-damage induced mutagenesis.
Our work is concerned with investigating DNA-damage induced mutagenesis, and the replication of damaged DNA which gives rise to these mutations, in budding yeast and E. coli. Such replication is used when unrepaired damage delays the progress of replication forks, and depends on a set of proteins at least partly different from those needed for replication on undamaged templates. Although only a minor cellular mechanism for coping with DNA damage, it is a major source of mutations. Understanding this process may therefore provide novel strategies for preventing cancer, since mutations commonly contribute to the development of this disease. To this end, we have cloned and sequenced yeast genes concerned with mutagenesis and, in collaboration with David Hinkle (Biology Department) examined the enzymatic properties of their purified products. In particular, we are investigating the properties of a new DNA polymerase, Pol zeta, a complex formed by the products of the REV3 and REV7 genes. The sole function of Pol zeta appears to be replication past damage sites in the DNA template, which it performs more efficiently than the major replicases. We are also investigating the properties of the REV1 gene product, which possesses a novel deoxycytidyl transferase activity that is required for replication past abasic lesions.
Studies with E. coli. carried out in collaboration with Roger Woodgate (NIH) and Myron Goodman (USC), are concerned with understanding the in vivo functions of the UmuD'C protein complex, which plays a crucial role in replication past DNA damage in this organism. In particular, we are attempting to identify the DNA polymerase holoenzyme subunits with which the complex interacts, and to determine the relative roles of DNA polymerase II and III. These questions are being investigated by transforming different mutant strains with single stranded vectors that carry a specified mutagenic lesion at a defined site. Such vectors are powerful tools for this purpose because they provide direct and independent estimates of the frequency of replication past the lesion and of the frequency with which mutations are induced by this process.
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Lawrence CW (2007) Following the RAD6 pathway. DNA Repair (Amst), 6:676-86
Zhang H, Lawrence CW (2005) The error-free component of the RAD6/RAD18 DNA damage tolerance pathway of budding yeast employs sister-strand recombination. Proc Natl Acad Sci U S A, 102:15954-9
Ozgenc AI, Szekeres ES, Lawrence CW (2005) In vivo evidence for a recA-independent recombination process in Escherichia coli that permits completion of replication of DNA containing UV damage in both strands. J Bacteriol, 187:1974-84
Gibbs PE, McDonald J, Woodgate R, Lawrence CW (2005) The relative roles in vivo of Saccharomyces cerevisiae Pol eta, Pol zeta, Rev1 protein and Pol32 in the bypass and mutation induction of an abasic site, T-T (6-4) photoadduct and T-T cis-syn cyclobutane dimer. Genetics, 169:575-82
Hoffman PD, Wang H, Lawrence CW, Iwai S, Hanaoka F, Hays JB (2005) Binding of MutS mismatch repair protein to DNA containing UV photoproducts, "mismatched" opposite Watson--Crick and novel nucleotides, in different DNA sequence contexts. DNA Repair (Amst), 4:983-93
Borden A, O'Grady PI, Vandewiele D, Fernandez de Henestrosa AR, Lawrence CW, Woodgate R (2002) Escherichia coli DNA polymerase III can replicate efficiently past a T-T cis-syn cyclobutane dimer if DNA polymerase V and the 3' to 5' exonuclease proofreading function encoded by dnaQ are inactivated. J Bacteriol, 184:2674-81
Lawrence CW (2002) Cellular roles of DNA polymerase zeta and Rev1 protein. DNA Repair (Amst), 1:425-35
Li Z, Zhang H, McManus TP, McCormick JJ, Lawrence CW, Maher VM (2002) hREV3 is essential for error-prone translesion synthesis past UV or benzo[a]pyrene diol epoxide-induced DNA lesions in human fibroblasts. Mutat Res, 510:71-80
Lawrence CW (2002) Classical mutagenesis techniques. Methods Enzymol, 350:189-99
Burgers PM, Koonin EV, Bruford E, Blanco L, Burtis KC, Christman MF, Copeland WC, Friedberg EC, Hanaoka F, Hinkle DC, Lawrence CW, Nakanishi M, Ohmori H, Prakash L, Prakash S, Reynaud CA, Sugino A, Todo T, Wang Z, Weill JC, Woodgate R (2001) Eukaryotic DNA polymerases: proposal for a revised nomenclature. J Biol Chem, 276:43487-90
Lawrence CW, Maher VM (2001) Eukaryotic mutagenesis and translesion replication dependent on DNA polymerase zeta and Rev1 protein. Biochem Soc Trans, 29:187-91
Ohmori H, Friedberg EC, Fuchs RP, Goodman MF, Hanaoka F, Hinkle D, Kunkel TA, Lawrence CW, Livneh Z, Nohmi T, Prakash L, Prakash S, Todo T, Walker GC, Wang Z, Woodgate R (2001) The Y-family of DNA polymerases. Mol Cell, 8:7-8
Lawrence CW, Maher VM (2001) Mutagenesis in eukaryotes dependent on DNA polymerase zeta and Rev1p. Philos Trans R Soc Lond B Biol Sci, 356:41-6
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E-Mail: Christopher_Lawrence@urmc.rochester.edu
Christopher Lawrence
Department of Biochemistry and Biophysics
University of Rochester School of Medicine and Dentistry
601 Elmwood Ave, Box 712
Rochester, New York 14642
Office: Medical Center Annex C-13
Telephone: (585) 275-2948; Fax: (585) 275-6007
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