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PMID: 8709248 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of latent membrane protein 2A (LMP2A) domains essential for the LMP2A dominant-negative effect on B-lymphocyte surface immunoglobulin signal transduction.

Journal of virology ·Vol. 70 ·No. 9 ·1996-09-00 ·Pages 6216-26

Fruehling S, Lee SK, Herrold R, Frech B, Laux G, Kremmer E, Grässer FA, Longnecker R

Abstract

Epstein-Barr virus (EBV) recombinants which carry three different deletion mutations in the LMP2A cytoplasmic amino-terminal domain were constructed. The presence of each mutation, LMP2A delta 21-36, LMP2A delta 21-64, and LMP2A delta 21-85, in EBV-infected transformed lymphoblastoid cell lines was confirmed by PCR analysis and Southern blot hybridization. Confirmation of mutant LMP2A protein expression was by immunofluorescence and immunoblotting with a newly identified rat monoclonal antibody that recognizes each of the LMP2A deletion mutations. Lymphoblastoid cell lines infected with recombinant EBV DNAs containing the mutations were analyzed for loss of LMP2A's dominant-negative effect on surface immunoglobulin signal transduction by monitoring induction of tyrosine phosphorylation, calcium mobilization, and activation of lytic replication following surface immunoglobulin cross-linking. Domains of LMP2A important for induction of tyrosine phosphorylation, calcium mobilization, and activation of lytic replication were identified.

MeSH Terms
Amino Acid Sequence Animals Antibodies, Monoclonal B-Lymphocytes/immunology Base Sequence Calcium/metabolism Cell Line, Transformed Cell Transformation, Viral DNA Primers DNA, Viral Fluorescent Antibody Technique Genetic Vectors Herpesvirus 4, Human/genetics,immunology,physiology Humans Molecular Sequence Data Mutagenesis Oncogene Proteins, Viral/physiology Phosphotyrosine/analysis Polymerase Chain Reaction Protein Structure, Secondary Rats Receptors, Antigen, B-Cell/physiology Recombination, Genetic Sequence Deletion Signal Transduction Viral Matrix Proteins/biosynthesis,chemistry,physiology Virus Replication
Chemicals
Antibodies, Monoclonal DNA Primers DNA, Viral EBV-associated membrane antigen, Epstein-Barr virus Oncogene Proteins, Viral Receptors, Antigen, B-Cell Viral Matrix Proteins Phosphotyrosine Calcium
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Fruehling S
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois, USA.
Lee S K
Herrold R
Frech B
Laux G
Kremmer E
Grässer F A
Longnecker R
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.70.9.6216-6226.1996
Published
1996-09-00
Pages
6216-26
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC190646
Subset
IM
Grants
NCI NIH HHS · CA62234 · United States
External Links
PubMed source DOI: 10.1128/JVI.70.9.6216-6226.1996 PMC Full Text View journal details
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