title: Pin1 interacts with the Epstein-Barr virus DNA polymerase catalytic subunit and regulates viral DNA replication.
abstract: Peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) protein is known as a regulator which recognizes phosphorylated Ser/Thr-Pro motifs and increases the rate of cis and trans amide isomer interconversion, thereby altering the conformation of its substrates. We found that Pin1 knockdown using short hairpin RNA (shRNA) technology resulted in strong suppression of productive Epstein-Barr virus (EBV) DNA replication. We further identified the EBV DNA polymerase catalytic subunit, BALF5, as a Pin1 substrate in glutathione S-transferase (GST) pulldown and immunoprecipitation assays. Lambda protein phosphatase treatment abolished the binding of BALF5 to Pin1, and mutation analysis of BALF5 revealed that replacement of the Thr178 residue by Ala (BALF5 T178A) disrupted the interaction with Pin1. To further test the effects of Pin1 in the context of virus infection, we constructed a BALF5-deficient recombinant virus. Exogenous supply of wild-type BALF5 in HEK293 cells with knockout recombinant EBV allowed efficient synthesis of viral genome DNA, but BALF5 T178A could not provide support as efficiently as wild-type BALF5. In conclusion, we found that EBV DNA polymerase BALF5 subunit interacts with Pin1 through BALF5 Thr178 in a phosphorylation-dependent manner. Pin1 might modulate EBV DNA polymerase conformation for efficient, productive viral DNA replication.
authors.affiliation: Division of Virology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Narita
authors.forename: Takayuki
authors.initials: T
authors.lastname: Murata
authors.forename: Akihide
authors.initials: A
authors.lastname: Ryo
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kawashima
authors.forename: Atsuko
authors.initials: A
authors.lastname: Sugimoto
authors.forename: Teru
authors.initials: T
authors.lastname: Kanda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Tsurumi
mesh_terms: D002460:Cell Line; D002498:Centrifugation; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000096:biosynthesis*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D004854:Herpesvirus 4, Human / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D025941:Protein Interaction Mapping; D013912:Threonine / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D014764:Viral Proteins; D013912:Threonine; C083202:BALF5 protein, Epstein-barr virus; D004259:DNA-Directed DNA Polymerase; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase