A novel mechanism for regulating the activity of proliferating cell nuclear antigen by a small protein Journal Article
| Authors: | Li, Z.; Huang, R. Y. C.; Yopp, D. C.; Hileman, T. H.; Santangelo, T. J.; Hurwitz, J.; Hudgens, J. W.; Kelman, Z. |
| Article Title: | A novel mechanism for regulating the activity of proliferating cell nuclear antigen by a small protein |
| Abstract: | Proliferating cell nuclear antigen (PCNA) forms a trimeric ring that associates with and influences the activity of many proteins participating in DNA metabolic processes and cell cycle progression. Previously, an uncharacterized small protein, encoded by TK0808 in the archaeon Thermococcus kodakarensis, was shown to stably interact with PCNA in vivo. Here, we show that this protein, designated Thermococcales inhibitor of PCNA (TIP), binds to PCNA in vitro and inhibits PCNA-dependent activities likely by preventing PCNA trimerization. Using hydrogen/deuterium exchange mass spectrometry and site-directed mutagenesis, the interacting regions of PCNA and TIP were identified. Most proteins bind to PCNA via a PCNA-interacting peptide (PIP) motif that interacts with the inter domain connecting loop (IDCL) on PCNA. TIP, however, lacks any known PCNA-interacting motif, suggesting a new mechanism for PCNA binding and regulation of PCNA-dependent activities, which may support the development of a new subclass of therapeutic biomolecules for inhibiting PCNA. © 2014 Published by Oxford University Press on behalf of Nucleic Acids Research 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US. |
| Keywords: | controlled study; unclassified drug; nonhuman; protein motif; ultraviolet radiation; mass spectrometry; protein binding; in vitro study; bacterial strain; molecular cloning; regulatory mechanism; protein purification; escherichia coli; polymerization; cycline; bacterial genome; mitomycin; site directed mutagenesis; protein inhibitor; bacterial growth; southern blotting; mesylic acid methyl ester; thermococcus kodakarensis; priority journal; article; gel permeation chromatography; thermococcales inhibitor of proliferating cell nuclear antigen; deuterium hydrogen exchange; methanothermobacter thermautotrophicus; trimerization |
| Journal Title: | Nucleic Acids Research |
| Volume: | 42 |
| Issue: | 9 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2014-01-01 |
| Start Page: | 5776 |
| End Page: | 5789 |
| Language: | English |
| DOI: | 10.1093/nar/gku239 |
| PROVIDER: | scopus |
| PMCID: | PMC4027161 |
| PUBMED: | 24728986 |
| DOI/URL: | |
| Notes: | Nucleic Acids Res. -- Export Date: 8 July 2014 -- CODEN: NARHA -- Source: Scopus |
