A cis element between the TATA box and the transcription start site of the major immediate-early promoter of human cytomegalovirus determines efficiency of viral replication

Hiroki Isomura, Mark F. Stinski, Ayumi Kudoh, Sanae Nakayama, Takayuki Murata, Yoshitaka Sato, Satoko Iwahori, Tatsuya Tsurumi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The promoter of the major immediate-early (MIE) genes of human cytomegalovirus (HCMV), also referred to as the CMV promoter, possesses a cis-acting element positioned downstream of the TATA box between positions -14 and -1 relative to the transcription start site (+1). We determined the role of the cis-acting element in viral replication by comparing recombinant viruses with the cis-acting element replaced with other sequences. Recombinant virus with the simian CMV counterpart replicated efficiently in human foreskin fibroblasts, as well as wild-type virus. In contrast, replacement with the murine CMV counterpart caused inefficient MIE gene transcription, RNA splicing, MIE and early viral gene expression, and viral DNA replication. To determine which nucleotides in the cis-acting element are required for efficient MIE gene transcription and splicing, we constructed mutations within the cis-acting element in the context of a recombinant virus. While mutations in the cis-acting element have only a minor effect on in vitro transcription, the effects on viral replication are major. The nucleotides at -10 and -9 in the cis-acting element relative to the transcription start site (+1) affect efficient MIE gene transcription and splicing at early times after infection. The cis-acting element also acts as a cis-repression sequence when the viral IE86 protein accumulates in the infected cell. We demonstrate that the cis-acting element has an essential role in viral replication.

Original languageEnglish
Pages (from-to)849-858
Number of pages10
JournalJournal of Virology
Volume82
Issue number2
DOIs
Publication statusPublished - 01-01-2008

Fingerprint

Human herpesvirus 5
TATA box
TATA Box
Immediate-Early Genes
Transcription Initiation Site
Genetic Transcription
virus replication
Cytomegalovirus
transcription (genetics)
promoter regions
Viruses
viruses
Nucleotides
RNA Splicing
Foreskin
Mutation
Viral Genes
Viral DNA
Viral Proteins
genes

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Isomura, Hiroki ; Stinski, Mark F. ; Kudoh, Ayumi ; Nakayama, Sanae ; Murata, Takayuki ; Sato, Yoshitaka ; Iwahori, Satoko ; Tsurumi, Tatsuya. / A cis element between the TATA box and the transcription start site of the major immediate-early promoter of human cytomegalovirus determines efficiency of viral replication. In: Journal of Virology. 2008 ; Vol. 82, No. 2. pp. 849-858.
@article{997388d71fd546b1b5d51cbb3854c22f,
title = "A cis element between the TATA box and the transcription start site of the major immediate-early promoter of human cytomegalovirus determines efficiency of viral replication",
abstract = "The promoter of the major immediate-early (MIE) genes of human cytomegalovirus (HCMV), also referred to as the CMV promoter, possesses a cis-acting element positioned downstream of the TATA box between positions -14 and -1 relative to the transcription start site (+1). We determined the role of the cis-acting element in viral replication by comparing recombinant viruses with the cis-acting element replaced with other sequences. Recombinant virus with the simian CMV counterpart replicated efficiently in human foreskin fibroblasts, as well as wild-type virus. In contrast, replacement with the murine CMV counterpart caused inefficient MIE gene transcription, RNA splicing, MIE and early viral gene expression, and viral DNA replication. To determine which nucleotides in the cis-acting element are required for efficient MIE gene transcription and splicing, we constructed mutations within the cis-acting element in the context of a recombinant virus. While mutations in the cis-acting element have only a minor effect on in vitro transcription, the effects on viral replication are major. The nucleotides at -10 and -9 in the cis-acting element relative to the transcription start site (+1) affect efficient MIE gene transcription and splicing at early times after infection. The cis-acting element also acts as a cis-repression sequence when the viral IE86 protein accumulates in the infected cell. We demonstrate that the cis-acting element has an essential role in viral replication.",
author = "Hiroki Isomura and Stinski, {Mark F.} and Ayumi Kudoh and Sanae Nakayama and Takayuki Murata and Yoshitaka Sato and Satoko Iwahori and Tatsuya Tsurumi",
year = "2008",
month = "1",
day = "1",
doi = "10.1128/JVI.01593-07",
language = "English",
volume = "82",
pages = "849--858",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "2",

}

A cis element between the TATA box and the transcription start site of the major immediate-early promoter of human cytomegalovirus determines efficiency of viral replication. / Isomura, Hiroki; Stinski, Mark F.; Kudoh, Ayumi; Nakayama, Sanae; Murata, Takayuki; Sato, Yoshitaka; Iwahori, Satoko; Tsurumi, Tatsuya.

In: Journal of Virology, Vol. 82, No. 2, 01.01.2008, p. 849-858.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A cis element between the TATA box and the transcription start site of the major immediate-early promoter of human cytomegalovirus determines efficiency of viral replication

AU - Isomura, Hiroki

AU - Stinski, Mark F.

AU - Kudoh, Ayumi

AU - Nakayama, Sanae

AU - Murata, Takayuki

AU - Sato, Yoshitaka

AU - Iwahori, Satoko

AU - Tsurumi, Tatsuya

PY - 2008/1/1

Y1 - 2008/1/1

N2 - The promoter of the major immediate-early (MIE) genes of human cytomegalovirus (HCMV), also referred to as the CMV promoter, possesses a cis-acting element positioned downstream of the TATA box between positions -14 and -1 relative to the transcription start site (+1). We determined the role of the cis-acting element in viral replication by comparing recombinant viruses with the cis-acting element replaced with other sequences. Recombinant virus with the simian CMV counterpart replicated efficiently in human foreskin fibroblasts, as well as wild-type virus. In contrast, replacement with the murine CMV counterpart caused inefficient MIE gene transcription, RNA splicing, MIE and early viral gene expression, and viral DNA replication. To determine which nucleotides in the cis-acting element are required for efficient MIE gene transcription and splicing, we constructed mutations within the cis-acting element in the context of a recombinant virus. While mutations in the cis-acting element have only a minor effect on in vitro transcription, the effects on viral replication are major. The nucleotides at -10 and -9 in the cis-acting element relative to the transcription start site (+1) affect efficient MIE gene transcription and splicing at early times after infection. The cis-acting element also acts as a cis-repression sequence when the viral IE86 protein accumulates in the infected cell. We demonstrate that the cis-acting element has an essential role in viral replication.

AB - The promoter of the major immediate-early (MIE) genes of human cytomegalovirus (HCMV), also referred to as the CMV promoter, possesses a cis-acting element positioned downstream of the TATA box between positions -14 and -1 relative to the transcription start site (+1). We determined the role of the cis-acting element in viral replication by comparing recombinant viruses with the cis-acting element replaced with other sequences. Recombinant virus with the simian CMV counterpart replicated efficiently in human foreskin fibroblasts, as well as wild-type virus. In contrast, replacement with the murine CMV counterpart caused inefficient MIE gene transcription, RNA splicing, MIE and early viral gene expression, and viral DNA replication. To determine which nucleotides in the cis-acting element are required for efficient MIE gene transcription and splicing, we constructed mutations within the cis-acting element in the context of a recombinant virus. While mutations in the cis-acting element have only a minor effect on in vitro transcription, the effects on viral replication are major. The nucleotides at -10 and -9 in the cis-acting element relative to the transcription start site (+1) affect efficient MIE gene transcription and splicing at early times after infection. The cis-acting element also acts as a cis-repression sequence when the viral IE86 protein accumulates in the infected cell. We demonstrate that the cis-acting element has an essential role in viral replication.

UR - http://www.scopus.com/inward/record.url?scp=37849008194&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=37849008194&partnerID=8YFLogxK

U2 - 10.1128/JVI.01593-07

DO - 10.1128/JVI.01593-07

M3 - Article

C2 - 17989180

AN - SCOPUS:37849008194

VL - 82

SP - 849

EP - 858

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 2

ER -