The human cytomegalovirus UL45 gene product is a late, virion-associated protein and influences virus growth at low multiplicities of infection

Marco Patrone, Elena Percivalle, Massimiliano Secchi, Loretta Fiorina, Guido Pedrali-Noy, Monica Zoppé, Fausto Baldanti, Gabriele Hahn, Ulrich H. Koszinowski, Gabriele Milanesi, Andrea Gallina

Research output: Contribution to journalArticle

Abstract

Human cytomegalovirus (HCMV) encodes a protein related to the large (R1) subunit of ribonucleotide reductase (RR), but does not encode the corresponding small (R2) subunit. The R1 homologue, UL45, lacks many catalytic residues, and its impact on deoxyribonucleoticle (dNTP) production remains unknown. Here, UL45 is shown to accumulate at late stages of infection and to be a virion tegument protein. To study UL45 function in its genome context, UL45 was disrupted by transposon insertion. The UL45-knockout (UL45-KO) mutant exhibited a growth defect in fibroblasts at a low m.o.i. and also a cell-to-cell spread defect. This did not result from a reduced dNTP supply because dNTP pools were unchanged in resting cells infected with the mutant virus. Irrespective of UL45 expression, all cellular RR subunits - S-phase RR subunits, and the p53-dependent p53R2 - were induced by infection. p53R2 was targeted to the infected cell nucleus, suggesting that HCMV diverts a mechanism normally activated by DNA damage response. Cells infected with the UL45-KO mutant were moderately sensitized to Fas-induced apoptosis relative to those infected with the parental virus. Together with the report on the UL45-KO endotheliotropic HCMV mutant (Hahn et al., J Virol 76, 9551-9555, 2002), these data suggest that UL45 does not share the prominent antiapototic role attributed to the mouse cytomegalovirus homologue M45.

Original languageEnglish
Pages (from-to)3359-3370
Number of pages12
JournalJournal of General Virology
Volume84
Issue number12
DOIs
Publication statusPublished - Dec 2003

Fingerprint

Cytomegalovirus
Ribonucleotide Reductases
Virion
Viruses
Growth
Infection
Genes
Proteins
Muromegalovirus
Cell Nucleus
S Phase
DNA Damage
Fibroblasts
Genome
Apoptosis

ASJC Scopus subject areas

  • Virology
  • Immunology

Cite this

The human cytomegalovirus UL45 gene product is a late, virion-associated protein and influences virus growth at low multiplicities of infection. / Patrone, Marco; Percivalle, Elena; Secchi, Massimiliano; Fiorina, Loretta; Pedrali-Noy, Guido; Zoppé, Monica; Baldanti, Fausto; Hahn, Gabriele; Koszinowski, Ulrich H.; Milanesi, Gabriele; Gallina, Andrea.

In: Journal of General Virology, Vol. 84, No. 12, 12.2003, p. 3359-3370.

Research output: Contribution to journalArticle

Patrone, Marco ; Percivalle, Elena ; Secchi, Massimiliano ; Fiorina, Loretta ; Pedrali-Noy, Guido ; Zoppé, Monica ; Baldanti, Fausto ; Hahn, Gabriele ; Koszinowski, Ulrich H. ; Milanesi, Gabriele ; Gallina, Andrea. / The human cytomegalovirus UL45 gene product is a late, virion-associated protein and influences virus growth at low multiplicities of infection. In: Journal of General Virology. 2003 ; Vol. 84, No. 12. pp. 3359-3370.
@article{bd9f684779c7429eb4f5552fbdaffdc7,
title = "The human cytomegalovirus UL45 gene product is a late, virion-associated protein and influences virus growth at low multiplicities of infection",
abstract = "Human cytomegalovirus (HCMV) encodes a protein related to the large (R1) subunit of ribonucleotide reductase (RR), but does not encode the corresponding small (R2) subunit. The R1 homologue, UL45, lacks many catalytic residues, and its impact on deoxyribonucleoticle (dNTP) production remains unknown. Here, UL45 is shown to accumulate at late stages of infection and to be a virion tegument protein. To study UL45 function in its genome context, UL45 was disrupted by transposon insertion. The UL45-knockout (UL45-KO) mutant exhibited a growth defect in fibroblasts at a low m.o.i. and also a cell-to-cell spread defect. This did not result from a reduced dNTP supply because dNTP pools were unchanged in resting cells infected with the mutant virus. Irrespective of UL45 expression, all cellular RR subunits - S-phase RR subunits, and the p53-dependent p53R2 - were induced by infection. p53R2 was targeted to the infected cell nucleus, suggesting that HCMV diverts a mechanism normally activated by DNA damage response. Cells infected with the UL45-KO mutant were moderately sensitized to Fas-induced apoptosis relative to those infected with the parental virus. Together with the report on the UL45-KO endotheliotropic HCMV mutant (Hahn et al., J Virol 76, 9551-9555, 2002), these data suggest that UL45 does not share the prominent antiapototic role attributed to the mouse cytomegalovirus homologue M45.",
author = "Marco Patrone and Elena Percivalle and Massimiliano Secchi and Loretta Fiorina and Guido Pedrali-Noy and Monica Zopp{\'e} and Fausto Baldanti and Gabriele Hahn and Koszinowski, {Ulrich H.} and Gabriele Milanesi and Andrea Gallina",
year = "2003",
month = "12",
doi = "10.1099/vir.0.19452-0",
language = "English",
volume = "84",
pages = "3359--3370",
journal = "Journal of General Virology",
issn = "0022-1317",
publisher = "Society for General Microbiology",
number = "12",

}

TY - JOUR

T1 - The human cytomegalovirus UL45 gene product is a late, virion-associated protein and influences virus growth at low multiplicities of infection

AU - Patrone, Marco

AU - Percivalle, Elena

AU - Secchi, Massimiliano

AU - Fiorina, Loretta

AU - Pedrali-Noy, Guido

AU - Zoppé, Monica

AU - Baldanti, Fausto

AU - Hahn, Gabriele

AU - Koszinowski, Ulrich H.

AU - Milanesi, Gabriele

AU - Gallina, Andrea

PY - 2003/12

Y1 - 2003/12

N2 - Human cytomegalovirus (HCMV) encodes a protein related to the large (R1) subunit of ribonucleotide reductase (RR), but does not encode the corresponding small (R2) subunit. The R1 homologue, UL45, lacks many catalytic residues, and its impact on deoxyribonucleoticle (dNTP) production remains unknown. Here, UL45 is shown to accumulate at late stages of infection and to be a virion tegument protein. To study UL45 function in its genome context, UL45 was disrupted by transposon insertion. The UL45-knockout (UL45-KO) mutant exhibited a growth defect in fibroblasts at a low m.o.i. and also a cell-to-cell spread defect. This did not result from a reduced dNTP supply because dNTP pools were unchanged in resting cells infected with the mutant virus. Irrespective of UL45 expression, all cellular RR subunits - S-phase RR subunits, and the p53-dependent p53R2 - were induced by infection. p53R2 was targeted to the infected cell nucleus, suggesting that HCMV diverts a mechanism normally activated by DNA damage response. Cells infected with the UL45-KO mutant were moderately sensitized to Fas-induced apoptosis relative to those infected with the parental virus. Together with the report on the UL45-KO endotheliotropic HCMV mutant (Hahn et al., J Virol 76, 9551-9555, 2002), these data suggest that UL45 does not share the prominent antiapototic role attributed to the mouse cytomegalovirus homologue M45.

AB - Human cytomegalovirus (HCMV) encodes a protein related to the large (R1) subunit of ribonucleotide reductase (RR), but does not encode the corresponding small (R2) subunit. The R1 homologue, UL45, lacks many catalytic residues, and its impact on deoxyribonucleoticle (dNTP) production remains unknown. Here, UL45 is shown to accumulate at late stages of infection and to be a virion tegument protein. To study UL45 function in its genome context, UL45 was disrupted by transposon insertion. The UL45-knockout (UL45-KO) mutant exhibited a growth defect in fibroblasts at a low m.o.i. and also a cell-to-cell spread defect. This did not result from a reduced dNTP supply because dNTP pools were unchanged in resting cells infected with the mutant virus. Irrespective of UL45 expression, all cellular RR subunits - S-phase RR subunits, and the p53-dependent p53R2 - were induced by infection. p53R2 was targeted to the infected cell nucleus, suggesting that HCMV diverts a mechanism normally activated by DNA damage response. Cells infected with the UL45-KO mutant were moderately sensitized to Fas-induced apoptosis relative to those infected with the parental virus. Together with the report on the UL45-KO endotheliotropic HCMV mutant (Hahn et al., J Virol 76, 9551-9555, 2002), these data suggest that UL45 does not share the prominent antiapototic role attributed to the mouse cytomegalovirus homologue M45.

UR - http://www.scopus.com/inward/record.url?scp=10744221040&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10744221040&partnerID=8YFLogxK

U2 - 10.1099/vir.0.19452-0

DO - 10.1099/vir.0.19452-0

M3 - Article

C2 - 14645917

AN - SCOPUS:10744221040

VL - 84

SP - 3359

EP - 3370

JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

IS - 12

ER -