The influence of the Cdc27 subunit on the properties of the Schizosaccharomyces pombe DNA polymerase δ Journal Article
| Authors: | Bermudez, V. P.; MacNeill, S. A.; Tappin, I.; Hurwitz, J. |
| Article Title: | The influence of the Cdc27 subunit on the properties of the Schizosaccharomyces pombe DNA polymerase δ |
| Abstract: | Schizosaccharomyces pombe DNA polymerase (pol) δ contains four subunits, pol 3, Cdc1, Cdc27, and Cdm1. In this report, we examined the role of Cdc27 on the structure and activity of pol δ. We show that the four-subunit complex is monomeric in structure, in contrast to the previous report that it was a dimer (Zuo, S., Bermudez, V., Zhang, G., Kelman, Z., and Hurwitz, J. (2000) J. Biol. Chem. 275, 5153-5162). This discrepancy between the earlier and recent observations was traced to the marked asymmetric shape of Cdc27. Cdc27 contains two critical domains that govern its role in activating pol δ. The N-terminal region (amino acids (aa) 1-160) binds to Cdc1 and its extreme C-terminal end (aa 362-369) interacts with proliferating cell nuclear antigen (PCNA). Mutants of S. pombe pol δ, containing truncated Cdc27 derivatives deficient in binding to PCNA, supported DNA replication less processively than the wild-type complex. Fusion of a minimal PCNA-binding motif (aa 352-372) to C-terminally truncated Cdc27 derivatives restored processive DNA synthesis in vitro. In vivo, the introduction of these fused Cdc27 derivatives into cdc27Δ cells conferred viability. These data support the model in which Cdc27 plays an essential role in DNA replication by recruiting PCNA to the pol δ holoenzyme. |
| Keywords: | unclassified drug; gene mutation; mutation; nonhuman; dna replication; protein conformation; protein domain; protein motif; cell cycle protein; animal cell; cell cycle proteins; cell viability; carboxy terminal sequence; protein protein interaction; protein binding; protein interaction; structure activity relation; wild type; animalia; biosynthesis; dna; amino terminal sequence; antigens; dimerization; protein structure, tertiary; amino acids; cycline; protein subunit; structure analysis; protein structure, quaternary; models, chemical; amino acid motifs; chromatography, gel; schizosaccharomyces; schizosaccharomyces pombe proteins; electrophoresis, polyacrylamide gel; schizosaccharomyces pombe; proliferating cell nuclear antigen; derivatives; dna directed dna polymerase delta; genetic complementation test; insect cell; polymerase; monomers; dimers; dna polymerase iii; amino acid derivative; insecta; priority journal; article; protein cdc1; protein cdc27; protein cdm1; protein pol3 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 277 |
| Issue: | 39 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2002-09-27 |
| Start Page: | 36853 |
| End Page: | 36862 |
| Language: | English |
| DOI: | 10.1074/jbc.M202897200 |
| PUBMED: | 12124382 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 14 November 2014 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work