The principal goal of this research is to study the enzymology of replication of yeast chromosomal DNA. The work will involve two aspects in DNA replication. In addition to studying replication systems in cell extracts that are specific for yeast replication origins, special attention will be paid to the purification and exact characterization of the enzymes involved in the elongation stage of replication: i.e. the DNA polymerases, the primase and the single-stranded DNA binding protein(s). Previous reports on the yeast DNA polymerases have to be reexamined in view of the fact that these studies almost certainly have been performed on proteolytic fragments. In analogy with the E. coli DNA polymerase III holoenzyme, a search will be made for a yeast holoenzyme using specific assay systems. The yeast primase willbe purified and we will determine whether there are conditions which make the priming event specific for particular single-stranded yeast DNA sequences. These purified replication proteins will be used in assay systems that measure replication from yeast replication origins. Replication mutants will be studied in vitro in these several assay systems. The proposed study of eucaryotic replication will focus on yeast because, of all eucaryotic organisms, yeast is the one most amenable to biochemical and genetic manipulation. An intensive study of DNA replication and its regulation in yeast will undoubtedly be of significance in understanding cell growth and division in higher organisms.
| Burgers, Peter M J; Kunkel, Thomas A (2017) Eukaryotic DNA Replication Fork. Annu Rev Biochem 86:417-438 |
| Koc, Katrina N; Singh, Saurabh P; Stodola, Joseph L et al. (2016) Pif1 removes a Rap1-dependent barrier to the strand displacement activity of DNA polymerase ?. Nucleic Acids Res 44:3811-9 |
| Burgers, Peter M J; Gordenin, Dmitry; Kunkel, Thomas A (2016) Who Is Leading the Replication Fork, Pol ? or Pol ?? Mol Cell 61:492-493 |
| Stodola, Joseph L; Stith, Carrie M; Burgers, Peter M (2016) Proficient Replication of the Yeast Genome by a Viral DNA Polymerase. J Biol Chem 291:11698-705 |
| Stojkovi?, Gorazd; Makarova, Alena V; Wanrooij, Paulina H et al. (2016) Oxidative DNA damage stalls the human mitochondrial replisome. Sci Rep 6:28942 |
| Stodola, Joseph L; Burgers, Peter M (2016) Resolving individual steps of Okazaki-fragment maturation at a millisecond timescale. Nat Struct Mol Biol 23:402-8 |
| Kochenova, Olga V; Bezalel-Buch, Rachel; Tran, Phong et al. (2016) Yeast DNA polymerase ? maintains consistent activity and mutagenicity across a wide range of physiological dNTP concentrations. Nucleic Acids Res : |
| Cho, Jang-Eun; Huang, Shar-Yin N; Burgers, Peter M et al. (2016) Parallel analysis of ribonucleotide-dependent deletions produced by yeast Top1 in vitro and in vivo. Nucleic Acids Res 44:7714-21 |
| Williams, Jessica S; Clausen, Anders R; Lujan, Scott A et al. (2015) Evidence that processing of ribonucleotides in DNA by topoisomerase 1 is leading-strand specific. Nat Struct Mol Biol 22:291-7 |
| Wanrooij, Paulina H; Burgers, Peter M (2015) Yet another job for Dna2: Checkpoint activation. DNA Repair (Amst) 32:17-23 |
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