A conserved interaction between the replicative clamp loader and DNA ligase in eukaryotes: Implications for Okazaki fragment joining Journal Article
| Authors: | Levin, D. S.; Vijayakumar, S.; Liu, X.; Bermudez, V. P.; Hurwitz, J.; Tomkinson, A. E. |
| Article Title: | A conserved interaction between the replicative clamp loader and DNA ligase in eukaryotes: Implications for Okazaki fragment joining |
| Abstract: | The recruitment of DNA ligase I to replication foci and the efficient joining of Okazaki fragments is dependent on the interaction between DNA ligase I and proliferating cell nuclear antigen (PCNA). Although the PCNA sliding clamp tethers DNA ligase I to nicked duplex DNA circles, the interaction does not enhance DNA joining. This suggests that other factors may be involved in the joining of Okazaki fragments. In this study, we describe an association between replication factor C (RFC), the clamp loader, and DNA ligase I in human cell extracts. Subsequently, we demonstrate that there is a direct physical interaction between these proteins that involves both the N- and C-terminal domains of DNA ligase I, the N terminus of the large RFC subunit p140, and the p36 and p38 subunits of RFC. Although RFC inhibited DNA joining by DNA ligase I, the addition of PCNA alleviated inhibition by RFC. Notably, the effect of PCNA on ligation was dependent on the PCNA-binding site of DNA ligase I. Together, these results provide a molecular explanation for the key in vivo role of the DNA ligase I/PCNA interaction and suggest that the joining of Okazaki fragments is coordinated by pairwise interactions among RFC, PCNA, and DNA ligase I. |
| Keywords: | human cell; dna-binding proteins; dna synthesis; proteins; enzyme inhibition; carboxy terminal sequence; protein protein interaction; protein binding; transcription, genetic; hela cell; hela cells; time factors; dna; antigens; eukaryota; glutathione transferase; protein biosynthesis; binding site; protein structure, tertiary; binding energy; binding sites; replication factor c; cycline; okazaki fragment; replication protein c; polydeoxyribonucleotide synthase; tissue; electrophoresis, polyacrylamide gel; dna ligases; biotin; proliferating cell nuclear antigen; cells; fungal proteins; chromatography; humans; human; priority journal; article; dna circles; human cell extracts; pcna binding sites; proliferating cell nuclear antigen (pcna) |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 279 |
| Issue: | 53 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2004-12-31 |
| Start Page: | 55196 |
| End Page: | 55201 |
| Language: | English |
| DOI: | 10.1074/jbc.M409250200 |
| PROVIDER: | scopus |
| PUBMED: | 15502161 |
| DOI/URL: | |
| Notes: | J. Biol. Chem. -- Cited By (since 1996):26 -- Export Date: 16 June 2014 -- CODEN: JBCHA -- Source: Scopus |
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