Prelich, G., Tan, C. K., Kostura, M., Mathews, M. B., So, A. G., Downey, K. M., Stillman, B. (April 1987) Functional identity of proliferating cell nuclear antigen and a DNA polymerase-delta auxiliary protein. Nature, 326 (6112). pp. 517-20. ISSN 0028-0836 (Print)
Abstract
The mechanism of replication of the simian virus 40 (SV40) genome closely resembles that of cellular chromosomes, thereby providing an excellent model system for examining the enzymatic requirements for DNA replication. Only one viral gene product, the large tumour antigen (large-T antigen), is required for viral replication, so the majority of replication enzymes must be cellular. Indeed, a number of enzymatic activities associated with replication and the S phase of the cell cycle are induced upon SV40 infection. Cell-free extracts derived from human cells, when supplemented with immunopurified SV40 large-T antigen support efficient replication of plasmids that contain the SV40 origin of DNA replication. Using this system, a cellular protein of relative molecular mass 36,000 (Mr = 36K) that is required for the elongation stage of SV40 DNA replication in vitro has been purified and identified as a known cell-cycle regulated protein, alternatively called the proliferating cell nuclear antigen (PCNA) or cyclin. It was noticed that, in its physical characteristics, PCNA closely resembles a protein that regulates the activity of calf thymus DNA polymerase-delta. Here we show that PCNA and the polymerase-delta auxiliary protein have similar electrophoretic behaviour and are both recognized by anti-PCNA human autoantibodies. More importantly, both proteins are functionally equivalent; they stimulate SV40 DNA replication in vitro and increase the processivity of calf thymus DNA polymerase-delta. These results implicate a novel animal cell DNA polymerase, DNA polymerase-delta, in the elongation stage of replicative DNA synthesis in vitro.
| Item Type: | Paper |
|---|---|
| Additional Information: | |
| Uncontrolled Keywords: | Antigens, Viral Tumor Autoantibodies immunology Autoantigens Cell Line DNA Polymerase III DNA Replication DNA-Directed DNA Polymerase metabolism Humans Kinetics Lupus Erythematosus Systemic immunology Nucleoproteins immunology physiology Proliferating Cell Nuclear Antigen Simian virus 40 genetics |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > DNA polymerase |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | CSHL Librarian |
| Date: | 2 April 1987 |
| Date Deposited: | 02 Mar 2012 18:33 |
| Last Modified: | 20 Jun 2017 20:31 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25044 |
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