A complex consisting of human replication factor C p40, p37, and p36 subunits is a DNA-dependent ATPase and an intermediate in the assembly of the holoenzyme Journal Article
| Authors: | Cai, J.; Gibbs, E.; Uhlmann, F.; Phillips, B.; Yao, N.; O'Donnell, M.; Hurwitz, J. |
| Article Title: | A complex consisting of human replication factor C p40, p37, and p36 subunits is a DNA-dependent ATPase and an intermediate in the assembly of the holoenzyme |
| Abstract: | Human replication factor C (hRFC) is a multi-subunit protein complex capable of supporting proliferating cell nuclear antigen (PCNA)-dependent DNA synthesis by DNA polymerases δ and ε. The hRFC complex consists of five different subunits with apparent molecular masses of 140, 40, 38, 37, and 36 kDa. We have previously reported the expression of a three-subunit core complex, consisting of the p40, p37, and p36 subunits following coupled in vitro transcription-translation of the cDNAs encoding these proteins (Uhlmann, F., Cai, J., Flores-Rozas, H., Dean, F. B., Finkelstein, J., O'Donnell, M., and Hurwitz, J. (1996) Proc. Natl. Acad. Sci. U.S. A. 93, 6521-6526). Here we describe the isolation of a stable complex composed of the p40, p37, and p36 subunits of hRFC from baculovirus-infected insect cells. The purified p40·p37·p36 complex, like the five-subunit RFC, contained DNA-dependent ATPase activity that was stimulated by PCNA, preferentially bound to primed DNA templates, interacted with PCNA, and was capable of unloading PCNA from singly-nicked circular DNA. In contrast to the five-subunit RFC, the three-subunit core complex did not lead PCNA onto DNA. The p40·p37·p36 complex inhibited the elongation of primed DNA templates catalyzed by the DNA polymerase δ holoenzyme. Incubation of the p40·p37·p36 complex with the hRFC p140 and p38 subunits formed the five- subunit hRFC complex that supported PCNA-dependent DNA synthesis by DNA polymerase δ. |
| Keywords: | dna-binding proteins; nonhuman; dna polymerase; dna replication; dna synthesis; animals; cell division; gene expression; protein protein interaction; homeodomain proteins; enzyme activity; hela cells; dna; escherichia coli; spodoptera; virus infection; saccharomyces cerevisiae proteins; catalysis; cycline; replication protein c; dna binding; adenosine triphosphatase; repressor proteins; molecular weight; adenosine triphosphatases; proto-oncogene proteins c-bcl-2; dna helicases; complementary dna; proliferating cell nuclear antigen; dna-directed dna polymerase; dna template; insect; dna protein complex; baculovirus; dna polymerase iii; unidentified baculovirus; insecta; dna, circular; humans; priority journal; article |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 272 |
| Issue: | 30 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1997-07-25 |
| Start Page: | 18974 |
| End Page: | 18981 |
| Language: | English |
| DOI: | 10.1074/jbc.272.30.18974 |
| PUBMED: | 9228079 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work