Brand, S. R., Bernstein, R. M., Mathews, M. B. (October 1994) Trimeric Structure of Human Proliferating Cell Nuclear Antigen Implications for Enzymatic Function and Autoantibody Recognition. Journal of Immunology, 153 (7). pp. 3070-3078. ISSN 0022-1767
Abstract
The proliferating cell nuclear Ag (PCNA) is a DNA replication factor postulated to function as a sliding damp around DNA. PCNA is also a target for autoimmunity in systemic lupus erythematosus. The autoantigenicity of PCNA is highly conformation-dependent, and reaction with most anti-PCNA sera requires a nearly full-length PCNA molecule. Here we describe the use of gel filtration and glycerol gradient sedimentation to analyze the native structure and size of PCNA. PCNA from three sources was studied (PCNA from HeLa cells, PCNA purified after its overexpression in bacteria, and PCNA produced in the wheat germ cell-free translation system) as well as mutant forms of PCNA translated in vitro. In each case, full-length PCNA behaved as a trimer. Analysis of mutant proteins revealed a correlation between the trimeric form and binding to the common type of human anti-PCNA autoantibody, suggesting that the Abs are specific for the active form of the protein. These findings are consistent with the idea that autoantibodies are generated as a response to native Ag and provide experimental support for the hypothesis that PCNA serves its processive function in DNA replication as a trimeric ring structure.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | DNA-POLYMERASE-DELTA AUXILIARY PROTEIN REPLICATION INVITRO CYCLIN PCNA S-PHASE EPITOPES CLONING REPAIR GENE RNA |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > immunoglobulin proteins |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | October 1994 |
| Date Deposited: | 27 Aug 2015 21:08 |
| Last Modified: | 27 Aug 2015 21:08 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/31399 |
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