Vpr protein of human immunodeficiency virus type 1 binds to 14-3-3 proteins and facilitates complex formation with Cdc25C: Implications for cell cycle arrest

Tomoshige Kino, Alexander Gragerov, Antonio Valentin, Maria Tsopanomihalou, Galina Ilyina-Gragerova, Rebecca Erwin-Cohen, George P. Chrousos, George N. Pavlakis

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Vpr and selected mutants were used in a Saccharomyces cerevisiae two-hybrid screen to identify cellular interactors. We found Vpr interacted with 14-3-3 proteins, a family regulating a multitude of proteins in the cell. Vpr mutant R80A, which is inactive in cell cycle arrest, did not interact with 14-3-3. 14-3-3 proteins regulate the G2/M transition by inactivating Cdc25C phosphatase via binding to the phosphorylated serine residue at position 216 of Cdc25C. 14-3-3 overexpression in human cells synergized with Vpr in the arrest of cell cycle. Vpr did not arrest efficiently cells not expressing 14-3-3σ. This indicated that a full complement of 14-3-3 proteins is necessary for optimal Vpr function on the cell cycle. Mutational analysis showed that the C-terminal portion of Vpr, known to harbor its cell cycle-arresting activity, bound directly to the C-terminal part of 14-3-3, outside of its phosphopeptide-binding pocket. Vpr expression shifted localization of the mutant Cdc25C S216A to the cytoplasm, indicating that Vpr promotes the association of 14-3-3 and Cdc25C, independently of the presence of serine 216. Immunoprecipitations of cell extracts indicated the presence of triple complexes (Vpr/14-3-3/Cdc25C). These results indicate that Vpr promotes cell cycle arrest at the G2/M phase by facilitating association of 14-3-3 and Cdc25C independently of the latter's phosphorylation status.

Original languageEnglish
Pages (from-to)2780-2787
Number of pages8
JournalJournal of Virology
Volume79
Issue number5
DOIs
Publication statusPublished - Mar 2005
Externally publishedYes

Fingerprint

vpr Gene Products
14-3-3 Proteins
Cell Cycle Checkpoints
HIV-1
Serine
Cell Cycle
cdc25 Phosphatases
Phosphopeptides
Complement C3
G2 Phase
Cell Extracts
Immunoprecipitation
Cell Division
Saccharomyces cerevisiae
Cytoplasm
Phosphorylation
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Vpr protein of human immunodeficiency virus type 1 binds to 14-3-3 proteins and facilitates complex formation with Cdc25C : Implications for cell cycle arrest. / Kino, Tomoshige; Gragerov, Alexander; Valentin, Antonio; Tsopanomihalou, Maria; Ilyina-Gragerova, Galina; Erwin-Cohen, Rebecca; Chrousos, George P.; Pavlakis, George N.

In: Journal of Virology, Vol. 79, No. 5, 03.2005, p. 2780-2787.

Research output: Contribution to journalArticle

Kino, T, Gragerov, A, Valentin, A, Tsopanomihalou, M, Ilyina-Gragerova, G, Erwin-Cohen, R, Chrousos, GP & Pavlakis, GN 2005, 'Vpr protein of human immunodeficiency virus type 1 binds to 14-3-3 proteins and facilitates complex formation with Cdc25C: Implications for cell cycle arrest', Journal of Virology, vol. 79, no. 5, pp. 2780-2787. https://doi.org/10.1128/JVI.79.5.2780-2787.2005
Kino, Tomoshige ; Gragerov, Alexander ; Valentin, Antonio ; Tsopanomihalou, Maria ; Ilyina-Gragerova, Galina ; Erwin-Cohen, Rebecca ; Chrousos, George P. ; Pavlakis, George N. / Vpr protein of human immunodeficiency virus type 1 binds to 14-3-3 proteins and facilitates complex formation with Cdc25C : Implications for cell cycle arrest. In: Journal of Virology. 2005 ; Vol. 79, No. 5. pp. 2780-2787.
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AU - Tsopanomihalou, Maria

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