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.
authors.affiliation: Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, Rockville, MD 20850, USA.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Li
authors.affiliation: Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, Rockville, MD 20850, USA National Institute of Standards and Technology, 9600 Gudelsky Drive, Rockville, MD 20850, USA.
authors.forename: Richard Y-C
authors.initials: RY
authors.lastname: Huang
authors.affiliation: Department of Microbiology and Center for RNA Biology, Ohio State University, Columbus, OH 43210, USA.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Yopp
authors.affiliation: Department of Microbiology and Center for RNA Biology, Ohio State University, Columbus, OH 43210, USA.
authors.forename: Travis H
authors.initials: TH
authors.lastname: Hileman
authors.affiliation: Department of Microbiology and Center for RNA Biology, Ohio State University, Columbus, OH 43210, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Santangelo
authors.affiliation: Program of Molecular Biology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Jerard
authors.initials: J
authors.lastname: Hurwitz
authors.affiliation: Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, Rockville, MD 20850, USA National Institute of Standards and Technology, 9600 Gudelsky Drive, Rockville, MD 20850, USA.
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Hudgens
authors.affiliation: Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, Rockville, MD 20850, USA National Institute of Standards and Technology, 9600 Gudelsky Drive, Rockville, MD 20850, USA zkelman@umd.edu.
authors.forename: Zvi
authors.initials: Z
authors.lastname: Kelman
mesh_terms: D019943:Amino Acid Substitution; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics; D004257:DNA Polymerase II / Q000737:chemistry; D041961:Deuterium Exchange Measurement; D045585:Flap Endonucleases / Q000737:chemistry; D007700:Kinetics; D050296:Microbial Viability; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D019712:Thermococcus / Q000502:physiology
chemical_list: D019843:Archaeal Proteins; D018809:Proliferating Cell Nuclear Antigen; D004257:DNA Polymerase II; D045585:Flap Endonucleases
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