Evidence that the Human Cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts

Patrizia Caposio; Ludovica Riera; Gabriele Hahn; Santo Landolfo; Giorgio Gribaudo

doi:10.1016/j.virol.2004.05.010

Evidence that the Human Cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts

Patrizia Caposio
, Ludovica Riera
, Gabriele Hahn
, Santo Landolfo
, Giorgio Gribaudo

Research output: Contribution to journal › Article › peer-review

28 Scopus citations

Abstract

The Human Cytomegalovirus (HCMV) UL72 gene is considered to be the equivalent of the dUTPase gene of the Alpha- and Gamma-herpesviruses. To characterize its function, the expression profiles of UL72 at both the RNA and the protein level were determined. The gene is expressed with a late kinetics and the corresponding UL72 46-kDa protein accumulates late during infection in the cytoplasm of infected cells. The pUL72 was expressed in E. coli and the purified recombinant protein did not display a detectable dUTPase activity. The viral yields of reconstituted HCMV RVΔUL72 viruses carrying a deletion within the UL72 ORF demonstrated a moderate growth defect following low MOI infections, whereas their DNA synthesis profiles were not significantly different from those of the parental HCMV RVAD169. These results demonstrate that the UL72 gene product is not a dUTPase and is not essential for replication in human fibroblasts.

Original language	English (US)
Pages (from-to)	264-276
Number of pages	13
Journal	Virology
Volume	325
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2004.05.010
State	Published - Aug 1 2004
Externally published	Yes

Funding

PC and LR equally contributed to this work. We thank Tokuhiro Chano for providing the NdUTPase cDNA, Giuseppe Gerna for the VR5438 and VR6264 strains, and Chistian Sinzger for the TB40/E strain. This work was supported by grants from MURST (40% and 60%), from the AIDS Research Project, and from the Ricerca Sanitaria Finalizzata (Regione Piemonte).

Funders
AIDS Research Project
MURST

Keywords

Deletion mutant viruses
Human cytomegalovirus
UL72 gene
dUTPase

ASJC Scopus subject areas

Virology

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10.1016/j.virol.2004.05.010

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title = "Evidence that the Human Cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts",

abstract = "The Human Cytomegalovirus (HCMV) UL72 gene is considered to be the equivalent of the dUTPase gene of the Alpha- and Gamma-herpesviruses. To characterize its function, the expression profiles of UL72 at both the RNA and the protein level were determined. The gene is expressed with a late kinetics and the corresponding UL72 46-kDa protein accumulates late during infection in the cytoplasm of infected cells. The pUL72 was expressed in E. coli and the purified recombinant protein did not display a detectable dUTPase activity. The viral yields of reconstituted HCMV RVΔUL72 viruses carrying a deletion within the UL72 ORF demonstrated a moderate growth defect following low MOI infections, whereas their DNA synthesis profiles were not significantly different from those of the parental HCMV RVAD169. These results demonstrate that the UL72 gene product is not a dUTPase and is not essential for replication in human fibroblasts.",

keywords = "Deletion mutant viruses, Human cytomegalovirus, UL72 gene, dUTPase",

author = "Patrizia Caposio and Ludovica Riera and Gabriele Hahn and Santo Landolfo and Giorgio Gribaudo",

note = "Funding Information: PC and LR equally contributed to this work. We thank Tokuhiro Chano for providing the NdUTPase cDNA, Giuseppe Gerna for the VR5438 and VR6264 strains, and Chistian Sinzger for the TB40/E strain. This work was supported by grants from MURST (40\% and 60\%), from the AIDS Research Project, and from the Ricerca Sanitaria Finalizzata (Regione Piemonte).",

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doi = "10.1016/j.virol.2004.05.010",

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AU - Landolfo, Santo

AU - Gribaudo, Giorgio

N1 - Funding Information: PC and LR equally contributed to this work. We thank Tokuhiro Chano for providing the NdUTPase cDNA, Giuseppe Gerna for the VR5438 and VR6264 strains, and Chistian Sinzger for the TB40/E strain. This work was supported by grants from MURST (40% and 60%), from the AIDS Research Project, and from the Ricerca Sanitaria Finalizzata (Regione Piemonte).

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N2 - The Human Cytomegalovirus (HCMV) UL72 gene is considered to be the equivalent of the dUTPase gene of the Alpha- and Gamma-herpesviruses. To characterize its function, the expression profiles of UL72 at both the RNA and the protein level were determined. The gene is expressed with a late kinetics and the corresponding UL72 46-kDa protein accumulates late during infection in the cytoplasm of infected cells. The pUL72 was expressed in E. coli and the purified recombinant protein did not display a detectable dUTPase activity. The viral yields of reconstituted HCMV RVΔUL72 viruses carrying a deletion within the UL72 ORF demonstrated a moderate growth defect following low MOI infections, whereas their DNA synthesis profiles were not significantly different from those of the parental HCMV RVAD169. These results demonstrate that the UL72 gene product is not a dUTPase and is not essential for replication in human fibroblasts.

AB - The Human Cytomegalovirus (HCMV) UL72 gene is considered to be the equivalent of the dUTPase gene of the Alpha- and Gamma-herpesviruses. To characterize its function, the expression profiles of UL72 at both the RNA and the protein level were determined. The gene is expressed with a late kinetics and the corresponding UL72 46-kDa protein accumulates late during infection in the cytoplasm of infected cells. The pUL72 was expressed in E. coli and the purified recombinant protein did not display a detectable dUTPase activity. The viral yields of reconstituted HCMV RVΔUL72 viruses carrying a deletion within the UL72 ORF demonstrated a moderate growth defect following low MOI infections, whereas their DNA synthesis profiles were not significantly different from those of the parental HCMV RVAD169. These results demonstrate that the UL72 gene product is not a dUTPase and is not essential for replication in human fibroblasts.

KW - Deletion mutant viruses

KW - Human cytomegalovirus

KW - UL72 gene

KW - dUTPase

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Evidence that the Human Cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts

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