Phorbol Ester-Induced Human Cytomegalovirus Major Immediate-Early (MIE) Enhancer Activation through PKC-Delta, CREB, and NF-κB Desilences MIE Gene Expression in Quiescently Infected Human Pluripotent NTera2 Cells

Xiaoqiu Liu; Jinxiang Yuan; Allen W Wu; Patrick W McGonagill; Courtney S Galle; Jeffery L Meier

doi:10.1128/JVI.00416-10

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Phorbol Ester-Induced Human Cytomegalovirus Major Immediate-Early (MIE) Enhancer Activation through PKC-Delta, CREB, and NF-κB Desilences MIE Gene Expression in Quiescently Infected Human Pluripotent NTera2 Cells

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Phorbol Ester-Induced Human Cytomegalovirus Major Immediate-Early (MIE) Enhancer Activation through PKC-Delta, CREB, and NF-κB Desilences MIE Gene Expression in Quiescently Infected Human Pluripotent NTera2 Cells

Xiaoqiu Liu, Jinxiang Yuan, Allen W Wu, Patrick W McGonagill, Courtney S Galle and Jeffery L Meier

Journal of virology, Vol.84(17), pp.8495-8508

09/2010

DOI: 10.1128/JVI.00416-10

PMCID: PMC2919020

PMID: 20504934

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Abstract

The ways in which human cytomegalovirus (HCMV) major immediate-early (MIE) gene expression breaks silence from latency to initiate the viral replicative cycle are poorly understood. A delineation of the signaling cascades that desilence the HCMV MIE genes during viral quiescence in the human pluripotent N-Tera2 (NT2) cell model provides insight into the molecular mechanisms underlying HCMV reactivation. In this model, we show that phorbol 12-myristate 13-acetate (PMA) immediately activates the expression of HCMV MIE RNA and protein and greatly increases the MIE-positive (MIE + ) NT2 cell population density; levels of Oct4 (pluripotent cell marker) and HCMV genome penetration are unchanged. Decreasing PKC-delta activity (pharmacological, dominant-negative, or RNA interference [RNAi] method) attenuates PMA-activated MIE gene expression. MIE gene activation coincides with PKC-delta Thr505 phosphorylation. Mutations in MIE enhancer binding sites for either CREB (cyclic AMP [cAMP] response element [CRE]) or NF-κB (κB) partially block PMA-activated MIE gene expression; the ETS binding site is negligibly involved, and κB does not confer MIE gene activation by vasoactive intestinal peptide (VIP). The PMA response is also partially attenuated by the RNAi-mediated depletion of the CREB or NF-κB subunit RelA or p50; it is not diminished by TORC2 knockdown or accompanied by TORC2 dephosphorylation. Mutations in both CRE and κB fully abolish PMA-activated MIE gene expression. Thus, PMA stimulates a PKC-delta-dependent, TORC2-independent signaling cascade that acts through cellular CREB and NF-κB, as well as their cognate binding sites in the MIE enhancer, to immediately desilence HCMV MIE genes. This signaling cascade is distinctly different from that elicited by VIP.

Genome and Regulation of Viral Gene Expression

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Title: Subtitle: Phorbol Ester-Induced Human Cytomegalovirus Major Immediate-Early (MIE) Enhancer Activation through PKC-Delta, CREB, and NF-κB Desilences MIE Gene Expression in Quiescently Infected Human Pluripotent NTera2 Cells
Creators: Xiaoqiu Liu - Veterans Affairs Medical Center
Jinxiang Yuan - Veterans Affairs Medical Center
Allen W Wu - Veterans Affairs Medical Center
Patrick W McGonagill - Veterans Affairs Medical Center
Courtney S Galle - Veterans Affairs Medical Center
Jeffery L Meier - Veterans Affairs Medical Center
Resource Type: Journal article
Publication Details: Journal of virology, Vol.84(17), pp.8495-8508
DOI: 10.1128/JVI.00416-10
PMID: 20504934
PMCID: PMC2919020
NLM abbreviation: J Virol
ISSN: 0022-538X
eISSN: 1098-5514
Publisher: American Society for Microbiology (ASM)
Language: English
Date published: 09/2010
Academic Unit: Infectious Diseases; Epidemiology; Surgery; Internal Medicine
Record Identifier: 9984051565002771

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