A DNA-Unwinding Protein Isolated from Escherichia coli: Its Interaction with DNA and with DNA Polymerases |
| |
| Authors: | Nolan Sigal Hajo Delius Thomas Kornberg Malcolm L Gefter and Bruce Alberts |
| |
| Institution: | Department of Biochemical Sciences, Princeton University, Princeton, New Jersey 08540;*Cold Spring Harbor Laboratory, Cold Spring Harbor, Long Island, New York 11724;?Department of Biological Sciences, Columbia University, New York, N.Y. 10027 |
| |
| Abstract: | A DNA-unwinding protein has been purified to homogeneity from E. coli. This protein has a molecular weight of about 22,000, as judged by its electrophoretic mobility on polyacrylamide gels containing sodium dodecylsulfate, and it appears to be present in about 800 copies per log-phase cell. It binds tightly and cooperatively to single-stranded DNA, and much less tightly, if at all, to RNA or double-stranded DNA.Like the T4 gene-32 protein characterized previously, the E. coli DNA-unwinding protein depresses the melting temperature of double-stranded DNAs, with regions rich in A-T base-pairs being preferentially melted. The E. coli protein strongly stimulates in vitro DNA synthesis by E. coli DNA polymerase II on appropriate templates; however, no stimulation is found with purified polymerases I or III of E. coli, or with T4 DNA polymerase. In contrast, gene-32 protein stimulates only the T4 DNA polymerase in a parallel assay. |
| |
| Keywords: | |
|
|
