Restricted protein phosphatase 2A targeting by merkel cell polyomavirus small T antigen

Hyun Jin Kwun, Masahiro Shuda, Carlos J. Camacho, Armin M. Gamper, Mamie Thant, Yuan Chang, Patrick S. Moore

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Merkel cell polyomavirus (MCV) is a newly discovered human cancer virus encoding a small T (sT) oncoprotein. We performed MCV sT FLAG-affinity purification followed by mass spectroscopy (MS) analysis, which identified several protein phosphatases (PP), including PP2A A and C subunits and PP4C, as potential cellular interacting proteins. PP2A targeting is critical for the transforming properties of nonhuman polyomaviruses, such as simian virus 40 (SV40), but is not required for MCV sT-induced rodent cell transformation. We compared similarities and differences in PP2A binding between MCV and SV40 sT. While SV40 sT coimmunopurified with subunits PP2A Aα and PP2A C, MCV sT coimmunopurified with PP2A Aα, PP2A Aβ, and PP2A C. Scanning alanine mutagenesis at 29 sites across the MCV sT protein revealed that PP2A-binding domains lie on the opposite molecular surface from a previously described large T stabilization domain (LSD) loop that binds E3 ligases, such as Fbw7. MCV sT-PP2A interactions can be functionally distinguished by mutagenesis from MCV sT LSD-dependent 4E-BP1 hyperphosphorylation and viral DNA replication enhancement. MCV sT has a restricted range for PP2A B subunit substitution, inhibiting only the assembly of B56α into the phosphatase holoenzyme. In contrast, SV40 sT inhibits the assembly of B55α, B56α and B56ε into PP2A. We conclude that MCV sT is required for Merkel cell carcinoma growth, but its in vitro transforming activity depends on LSD interactions rather than PP2A targeting.

Original languageEnglish (US)
Pages (from-to)4191-4200
Number of pages10
JournalJournal of virology
Volume89
Issue number8
DOIs
StatePublished - Jan 1 2015

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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    Kwun, H. J., Shuda, M., Camacho, C. J., Gamper, A. M., Thant, M., Chang, Y., & Moore, P. S. (2015). Restricted protein phosphatase 2A targeting by merkel cell polyomavirus small T antigen. Journal of virology, 89(8), 4191-4200. https://doi.org/10.1128/JVI.00157-15