Ng, L., Prelich, G., Anderson, C. W., Stillman, B., Fisher, P. A. (July 1990) Drosophila Proliferating Cell Nuclear Antigen - Structural and Functional Homology with Its Mammalian Counterpart. Journal of Biological Chemistry, 265 (20). pp. 11948-11954. ISSN 0021-9258
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Abstract
A protein with an apparent mass of 36 kDa was purified from Drosophila melanogaster embryos using a protocol developed for the purification of proliferating cell nuclear antigen (PCNA) from human 293 cells. The Drosophila protein comigrated with human PCNA on one-dimensional sodium dodecyl sulfate-polyacrylamide gels and cross-reacted with monoclonal anti-rabbit PCNA antibodies. NH2-terminal amino acid sequence analysis revealed that the putative Drosophila PCNA was highly homologous to human PCNA. Of the first 22 amino acids, 16 were identical, and 4 of the remaining 6 were changed conservatively. Results of total amino acid analysis were also consistent with a high degree of similarity between Drosophila PCNA and human PCNA. Functional analysis using the reconstituted simian virus 40 in vitro DNA replication system demonstrated that Drosophila PCNA could substitute, albeit with reduced efficiency, for human PCNA in stimulating simian virus 40 DNA synthesis. Affinity-purified anti-Drosophila PCNA antibodies cross-reacted with human PCNA and were able to recognize specifically Drosophila PCNA both on crude homogenate immunoblots and by indirect immunofluorescence analysis of proliferating cells in larval tissues in situ. These antibodies thus promise to be useful probes for the study of cell proliferation in this rapidly developing organism.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence Animals Antibodies isolation & purification Autoantigens genetics DNA Replication Drosophila melanogaster genetics growth & development immunology Electrophoresis Polyacrylamide Gel Fluorescent Antibody Technique Humans Immunoblotting Molecular Sequence Data Nuclear Proteins genetics/immunology isolation & purification Plasmids Proliferating Cell Nuclear Antigen Sequence Homology Nucleic Acid |
| Subjects: | organism description > animal > insect > Drosophila |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | CSHL Librarian |
| Date: | July 1990 |
| Date Deposited: | 02 Mar 2012 19:06 |
| Last Modified: | 20 Jun 2017 20:15 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25032 |
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