Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein

A. Gallina, L. Simoncini, S. Garbelli, E. Percivalle, G. Pedrali-Noy, K. S. Lee, R. L. Erikson, B. Plachter, G. Gerna, G. Milanesi

Research output: Contribution to journalArticle

Abstract

Human cytomegalovirus (HCMV) pp65 protein is the major constituent of viral dense bodies but is dispensable for viral growth in vitro, pp65 copurifies with a S/T kinase activity and has been implicated in phosphorylation of HCMV IE1 immediate-early protein and its escape from major histocompatibility complex 1 presentation. Furthermore, the presence of pp65 correlates with a virion-associated kinase activity. To clarify the role of pp65, yeast two-hybrid system (THS) screening was performed to identify pp65 cellular partners. A total of 18 out of 48 yeast clones harboring cDNAs for putative pp65 binding proteins encoded the Polo-like kinase 1 (Plk1) C- terminal domain. Plk1 behaved as a bona fide pp65 partner in THS control crosses, and the interaction was confirmed by in vitro binding experiments. Endogenous Plk1 was coimmunoprecipitated with pp65 from transiently transfected COS7 cells. In infected fibroblasts, Plk1 was coimmunoprecipitated with pp65 at late infection stages. Furthermore, Plk1 was detected within wild-type HCMV particles but not within the particles of a pp65-negative mutant (RVAd65). The hydrophilic region of pp65 was phosphorylated in vitro by Plk1. These results suggest that one function of pp65 may be to capture a cell kinase, perhaps in order to alter its activity, nucleotide preference, substrate specificity, or subcellular localization to the advantage of HCMV.

Original languageEnglish
Pages (from-to)1468-1478
Number of pages11
JournalJournal of Virology
Volume73
Issue number2
Publication statusPublished - 1999

Fingerprint

Human herpesvirus 5
phosphotransferases (kinases)
Phosphotransferases
proteins
Cytomegalovirus
two hybrid system techniques
Two-Hybrid System Techniques
Substrate Specificity
Major Histocompatibility Complex
Virion
yeasts
polo-like kinase 1
cytomegalovirus matrix protein 65kDa
Carrier Proteins
Nucleotides
Complementary DNA
Clone Cells
Fibroblasts
Yeasts
Phosphorylation

ASJC Scopus subject areas

  • Immunology

Cite this

Gallina, A., Simoncini, L., Garbelli, S., Percivalle, E., Pedrali-Noy, G., Lee, K. S., ... Milanesi, G. (1999). Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein. Journal of Virology, 73(2), 1468-1478.

Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein. / Gallina, A.; Simoncini, L.; Garbelli, S.; Percivalle, E.; Pedrali-Noy, G.; Lee, K. S.; Erikson, R. L.; Plachter, B.; Gerna, G.; Milanesi, G.

In: Journal of Virology, Vol. 73, No. 2, 1999, p. 1468-1478.

Research output: Contribution to journalArticle

Gallina, A, Simoncini, L, Garbelli, S, Percivalle, E, Pedrali-Noy, G, Lee, KS, Erikson, RL, Plachter, B, Gerna, G & Milanesi, G 1999, 'Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein', Journal of Virology, vol. 73, no. 2, pp. 1468-1478.
Gallina A, Simoncini L, Garbelli S, Percivalle E, Pedrali-Noy G, Lee KS et al. Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein. Journal of Virology. 1999;73(2):1468-1478.
Gallina, A. ; Simoncini, L. ; Garbelli, S. ; Percivalle, E. ; Pedrali-Noy, G. ; Lee, K. S. ; Erikson, R. L. ; Plachter, B. ; Gerna, G. ; Milanesi, G. / Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein. In: Journal of Virology. 1999 ; Vol. 73, No. 2. pp. 1468-1478.
@article{5d6a8d268b824fb9b26d7a2ec1c001d0,
title = "Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein",
abstract = "Human cytomegalovirus (HCMV) pp65 protein is the major constituent of viral dense bodies but is dispensable for viral growth in vitro, pp65 copurifies with a S/T kinase activity and has been implicated in phosphorylation of HCMV IE1 immediate-early protein and its escape from major histocompatibility complex 1 presentation. Furthermore, the presence of pp65 correlates with a virion-associated kinase activity. To clarify the role of pp65, yeast two-hybrid system (THS) screening was performed to identify pp65 cellular partners. A total of 18 out of 48 yeast clones harboring cDNAs for putative pp65 binding proteins encoded the Polo-like kinase 1 (Plk1) C- terminal domain. Plk1 behaved as a bona fide pp65 partner in THS control crosses, and the interaction was confirmed by in vitro binding experiments. Endogenous Plk1 was coimmunoprecipitated with pp65 from transiently transfected COS7 cells. In infected fibroblasts, Plk1 was coimmunoprecipitated with pp65 at late infection stages. Furthermore, Plk1 was detected within wild-type HCMV particles but not within the particles of a pp65-negative mutant (RVAd65). The hydrophilic region of pp65 was phosphorylated in vitro by Plk1. These results suggest that one function of pp65 may be to capture a cell kinase, perhaps in order to alter its activity, nucleotide preference, substrate specificity, or subcellular localization to the advantage of HCMV.",
author = "A. Gallina and L. Simoncini and S. Garbelli and E. Percivalle and G. Pedrali-Noy and Lee, {K. S.} and Erikson, {R. L.} and B. Plachter and G. Gerna and G. Milanesi",
year = "1999",
language = "English",
volume = "73",
pages = "1468--1478",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "2",

}

TY - JOUR

T1 - Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein

AU - Gallina, A.

AU - Simoncini, L.

AU - Garbelli, S.

AU - Percivalle, E.

AU - Pedrali-Noy, G.

AU - Lee, K. S.

AU - Erikson, R. L.

AU - Plachter, B.

AU - Gerna, G.

AU - Milanesi, G.

PY - 1999

Y1 - 1999

N2 - Human cytomegalovirus (HCMV) pp65 protein is the major constituent of viral dense bodies but is dispensable for viral growth in vitro, pp65 copurifies with a S/T kinase activity and has been implicated in phosphorylation of HCMV IE1 immediate-early protein and its escape from major histocompatibility complex 1 presentation. Furthermore, the presence of pp65 correlates with a virion-associated kinase activity. To clarify the role of pp65, yeast two-hybrid system (THS) screening was performed to identify pp65 cellular partners. A total of 18 out of 48 yeast clones harboring cDNAs for putative pp65 binding proteins encoded the Polo-like kinase 1 (Plk1) C- terminal domain. Plk1 behaved as a bona fide pp65 partner in THS control crosses, and the interaction was confirmed by in vitro binding experiments. Endogenous Plk1 was coimmunoprecipitated with pp65 from transiently transfected COS7 cells. In infected fibroblasts, Plk1 was coimmunoprecipitated with pp65 at late infection stages. Furthermore, Plk1 was detected within wild-type HCMV particles but not within the particles of a pp65-negative mutant (RVAd65). The hydrophilic region of pp65 was phosphorylated in vitro by Plk1. These results suggest that one function of pp65 may be to capture a cell kinase, perhaps in order to alter its activity, nucleotide preference, substrate specificity, or subcellular localization to the advantage of HCMV.

AB - Human cytomegalovirus (HCMV) pp65 protein is the major constituent of viral dense bodies but is dispensable for viral growth in vitro, pp65 copurifies with a S/T kinase activity and has been implicated in phosphorylation of HCMV IE1 immediate-early protein and its escape from major histocompatibility complex 1 presentation. Furthermore, the presence of pp65 correlates with a virion-associated kinase activity. To clarify the role of pp65, yeast two-hybrid system (THS) screening was performed to identify pp65 cellular partners. A total of 18 out of 48 yeast clones harboring cDNAs for putative pp65 binding proteins encoded the Polo-like kinase 1 (Plk1) C- terminal domain. Plk1 behaved as a bona fide pp65 partner in THS control crosses, and the interaction was confirmed by in vitro binding experiments. Endogenous Plk1 was coimmunoprecipitated with pp65 from transiently transfected COS7 cells. In infected fibroblasts, Plk1 was coimmunoprecipitated with pp65 at late infection stages. Furthermore, Plk1 was detected within wild-type HCMV particles but not within the particles of a pp65-negative mutant (RVAd65). The hydrophilic region of pp65 was phosphorylated in vitro by Plk1. These results suggest that one function of pp65 may be to capture a cell kinase, perhaps in order to alter its activity, nucleotide preference, substrate specificity, or subcellular localization to the advantage of HCMV.

UR - http://www.scopus.com/inward/record.url?scp=0032902559&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032902559&partnerID=8YFLogxK

M3 - Article

C2 - 9882353

AN - SCOPUS:0032902559

VL - 73

SP - 1468

EP - 1478

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 2

ER -