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PMID: 9151869 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Epstein-Barr virus latent membrane protein 2 associates with and is a substrate for mitogen-activated protein kinase.

Journal of virology ·Vol. 71 ·No. 6 ·1997-06-00 ·Pages 4752-60

Panousis CG, Rowe DT

Abstract

The latent membrane protein 2 (LMP2) of Epstein-Barr virus interferes with B-lymphocyte signal transduction through the immunoglobulin (Ig) receptor. Two isoforms of LMP2 exist and differ only in that one isoform (LMP2a) contains an N-terminal cytoplasmic domain that the other isoform does not. LMP2a is a phosphoprotein that is phosphorylated on tyrosines and serines in the cytoplasmic domain. GST1-119, a glutathione S-transferase (GST) fusion protein containing the 119 amino acids of the cytoplasmic domain, affinity precipitated serine kinase activity from BJAB cell extracts. The affinity-precipitated kinase phosphorylated LMP2a sequences, and kinase activity was increased following induction. Probing of Western immunoblots of affinity-precipitated proteins showed that the Erk1 form of mitogen-activated protein kinase (MAPK) was present. Purified MAPK phosphorylated GST fusion proteins containing the cytoplasmic domain of LMP2a and mutational analyses were used to identify S15 and S102 as the sites of in vitro phosphorylation. A polyclonal rabbit antiserum was prepared against a maltose binding protein-LMP2a cytoplasmic domain fusion protein (MBP1-119) and used to immunoprecipitate LMP2a from the in vitro-immortalized lymphoblastoid B-cell line B95-8CR. LMP2a immunoprecipitates from B95-8CR contained MAPK as a coprecipitated protein. Cross-linking surface Ig on B95-8CR cells failed to induce MAPK activity within the cells. Treatment of B95-8CR with phorbol myristate acetate (PMA) was able to bypass the Ig receptor block and activate MAPK activity. Phosphorylation of LMP2a on serine residues increased after PMA induction. The possible role for LMP2a serine phosphorylation by MAPK in the control of latency is discussed.

MeSH Terms
Amino Acid Sequence Animals Calcium-Calmodulin-Dependent Protein Kinases/metabolism Enzyme Activation Enzyme Induction Extracellular Space Humans Immunologic Techniques Macromolecular Substances Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases Molecular Sequence Data Phosphorylation Phosphoserine/chemistry Protein Binding Substrate Specificity Viral Matrix Proteins/metabolism
Chemicals
EBV-associated membrane antigen, Epstein-Barr virus Macromolecular Substances Viral Matrix Proteins Phosphoserine Calcium-Calmodulin-Dependent Protein Kinases Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Panousis C G
Department of Infectious Diseases and Microbiology, Graduate School of Public Health, University of Pittsburgh, Pennsylvania 15261, USA.
Rowe D T
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.71.6.4752-4760.1997
Published
1997-06-00
Pages
4752-60
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC191697
Subset
IM
Grants
NCI NIH HHS · CA53560 · United States
External Links
PubMed source DOI: 10.1128/JVI.71.6.4752-4760.1997 PMC Full Text View journal details
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