Requirement of multiple cis-acting elements in the human cytomegalovirus major immediate-early distal enhancer for viral gene expression and replication

Jeffery L Meier; Michael J Keller; James J McCoy

doi:10.1128/JVI.76.1.313-326.2002

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Requirement of multiple cis-acting elements in the human cytomegalovirus major immediate-early distal enhancer for viral gene expression and replication

Journal article

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Peer reviewed

Requirement of multiple cis-acting elements in the human cytomegalovirus major immediate-early distal enhancer for viral gene expression and replication

Jeffery L Meier, Michael J Keller and James J McCoy

Journal of virology, Vol.76(1), pp.313-326

01/2002

DOI: 10.1128/JVI.76.1.313-326.2002

PMCID: PMC135711

PMID: 11739696

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Abstract

We have shown previously that the human cytomegalovirus (HCMV) major immediate-early (MIE) distal enhancer is needed for MIE promoter-dependent transcription and viral replication at low multiplicities of infection (MOI). To understand how this region works, we constructed and analyzed a series of HCMVs with various distal enhancer mutations. We show that the distal enhancer is composed of at least two parts that function independently to coordinately activate MIE promoter-dependent transcription and viral replication. One such part is contained in a 47-bp segment that has consensus binding sites for CREB/ATF, SP1, and YY1. At low MOI, these working parts likely function in cis to directly activate MIE gene expression, thus allowing viral replication to ensue. Three findings support the view that these working parts are likely cis-acting elements. (i) Deletion of either part of a bisegmented distal enhancer only slightly alters MIE gene transcription and viral replication. (ii) Reversing the distal enhancer's orientation largely preserves MIE gene transcription and viral replication. (iii) Placement of stop codons at -300 or -345 in all reading frames does not impair MIE gene transcription and viral replication. Lastly, we show that these working parts are dispensable at high MOI, partly because of compensatory stimulation of MIE promoter activity and viral replication that is induced by a virion-associated component(s) present at a high viral particle/cell ratio. We conclude that the distal enhancer is a complex multicomponent cis-acting region that is required to augment both MIE promoter-dependent transcription and HCMV replication.

Gene Expression

Immediate-Early Proteins - genetics

Immediate-Early Proteins - physiology

Virus Replication

Humans

Cells, Cultured

Transcription, Genetic

Enhancer Elements, Genetic - physiology

Mutation

Suppressor of Cytokine Signaling Proteins

Cytomegalovirus - physiology

Details

Title: Subtitle: Requirement of multiple cis-acting elements in the human cytomegalovirus major immediate-early distal enhancer for viral gene expression and replication
Creators: Jeffery L Meier - Department of Internal Medicine and the Helen C. Levitt Center for Viral Pathogenesis and Disease, University of Iowa College of Medicine, Iowa City, Iowa, 52242, USA. jeffery-meier@uiowa.edu
Michael J Keller
James J McCoy
Resource Type: Journal article
Publication Details: Journal of virology, Vol.76(1), pp.313-326
DOI: 10.1128/JVI.76.1.313-326.2002
PMID: 11739696
PMCID: PMC135711
NLM abbreviation: J Virol
ISSN: 0022-538X
eISSN: 1098-5514
Grant note: AI-40130 / NIAID NIH HHS
Language: English
Date published: 01/2002
Academic Unit: Infectious Diseases; Epidemiology; Internal Medicine
Record Identifier: 9984094493402771

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