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PMID: 9733815 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Tyrosine 112 of latent membrane protein 2A is essential for protein tyrosine kinase loading and regulation of Epstein-Barr virus latency.

Journal of virology ·Vol. 72 ·No. 10 ·1998-10-00 ·Pages 7796-806

Fruehling S, Swart R, Dolwick KM, Kremmer E, Longnecker R

Abstract

Latent membrane protein 2A (LMP2A) of Epstein-Barr virus (EBV) is expressed on the plasma membrane of B lymphocytes latently infected with EBV and blocks B-cell receptor (BCR) signal transduction in EBV-immortalized B cells in vitro. The LMP2A amino-terminal domain that is essential for the LMP2A-mediated block on BCR signal transduction contains eight tyrosine residues. Association of Syk protein tyrosine kinase (PTK) with LMP2A occurs at the two tyrosines of the LMP2A immunoreceptor tyrosine-based activation motif, and it is hypothesized that Lyn PTK associates with the YEEA amino acid motif at LMP2A tyrosine 112 (Y112). To examine the specific association of Lyn PTK to LMP2A, a panel of LMP2A cDNA expression vectors containing LMP2A mutations were transfected into an EBV-negative B-cell line and analyzed for Lyn and LMP2A coimmunoprecipitation. Lyn associates with wild-type LMP2A and other LMP2A mutant constructs, but Lyn association is lost in the LMP2A construct containing a tyrosine (Y)-to-phenylalanine (F) mutation at LMP2A residue Y112 (LMP2AY112F). Next, the LMP2AY112F mutation was recombined into the EBV genome to generate stable lymphoblastoid cell lines (LCLs) transformed with the LMP2AY112F mutant virus. Analysis of BCR-mediated signal transduction in the LMP2AY112F LCLs revealed loss of the LMP2A-mediated block in BCR signal transduction. In addition, LMP2A was not tyrosine phosphorylated in LMP2AY112F LCLs. Together these data indicate the importance of the LMP2A Y112 residue in the ability of LMP2A to block BCR-mediated signal transduction and place the role of this residue and its interaction with Lyn PTK as essential to LMP2A phosphorylation, PTK loading, and down-modulation of PTKs involved in BCR-mediated signal transduction.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Calcium/metabolism Cell Line DNA Primers DNA, Complementary DNA-Binding Proteins/genetics Herpesvirus 4, Human/physiology Humans Molecular Sequence Data Mutagenesis, Site-Directed Phosphorylation Protein-Tyrosine Kinases/metabolism Trans-Activators/genetics Tumor Cells, Cultured Tyrosine/metabolism Viral Matrix Proteins/chemistry,genetics,metabolism Viral Proteins Virus Latency
Chemicals
BZLF1 protein, Herpesvirus 4, Human DNA Primers DNA, Complementary DNA-Binding Proteins EBV-associated membrane antigen, Epstein-Barr virus Trans-Activators Viral Matrix Proteins Viral Proteins Tyrosine Protein-Tyrosine Kinases Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fruehling S
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Swart R
Dolwick K M
Kremmer E
Longnecker R
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.72.10.7796-7806.1998
Published
1998-10-00
Pages
7796-806
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC110092
Subset
IM
Grants
NCI NIH HHS · R01 CA062234 · United States
NCI NIH HHS · R01 CA073507 · United States
NCI NIH HHS · CA62234 · United States
NCI NIH HHS · CA73507 · United States
External Links
PubMed source DOI: 10.1128/JVI.72.10.7796-7806.1998 PMC Full Text View journal details
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