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PMID: 11024139 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interaction of Epstein-Barr virus nuclear antigen leader protein (EBNA-LP) with HS1-associated protein X-1: implication of cytoplasmic function of EBNA-LP.

Journal of virology ·Vol. 74 ·No. 21 ·2000-11-00 ·Pages 10104-11

Kawaguchi Y, Nakajima K, Igarashi M, Morita T, Tanaka M, Suzuki M, Yokoyama A, Matsuda G, Kato K, Kanamori M, Hirai K

Abstract

Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) consists of W1W2 repeats and a unique C-terminal Y1Y2 domain and has been suggested to play an important role in EBV-induced transformation. To identify the cellular factors interacting with EBNA-LP, we performed a yeast two-hybrid screen, using EBNA-LP cDNA containing four W1W2 repeats as bait and an EBV-transformed human peripheral blood lymphocyte cDNA library as the source of cellular genes. Our results were as follows. (i) All three cDNAs in positive yeast colonies were found to encode the same cellular protein, HS1-associated protein X-1 (HAX-1), which is localized mainly in the cytoplasm and has been suggested to be involved in the regulation of B-cell signal transduction and apoptosis. (ii) Mutational analysis of EBNA-LP revealed that the association with HAX-1 is mediated by the W1W2 repeat domain. (iii) A purified chimeric protein consisting of glutathione S-transferase fused to EBNA-LP specifically formed complexes with HAX-1 transiently expressed in COS-7 cells. (iv) When EBNA-LP and HAX-1 were coexpressed in COS-7 cells, EBNA-LP was specifically coimmunoprecipitated with HAX-1. (v) Careful cell fractionation experiments of an EBV-infected lymphoblastoid cell line revealed that EBNA-LP is localized in the cytoplasm as well as in the nucleus. (vi) When EBNA-LP containing four W1W2 repeats was expressed in COS-7 cells, EBNA-LP was detected mainly in the nucleus by immunofluorescence assay. Interestingly, when EBNA-LP containing a single W1W2 repeat was expressed in COS-7 cells, EBNA-LP was localized predominantly in the cytoplasm and was colocalized with HAX-1. These results indicate that EBNA-LP is in fact present and may have a significant function in the cytoplasm, possibly by interacting with and affecting the function of HAX-1.

MeSH Terms
Adaptor Proteins, Signal Transducing Animals COS Cells Cell Line Cell Transformation, Viral Cytoplasm/metabolism Herpesvirus 4, Human/genetics,metabolism Humans Immunoblotting Plasmids/genetics Precipitin Tests Proteins/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism Transfection Two-Hybrid System Techniques Viral Proteins/genetics,metabolism
Chemicals
Adaptor Proteins, Signal Transducing EBNA-LP protein, Human herpesvirus 4 HAX1 protein, human Proteins Recombinant Fusion Proteins Viral Proteins
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Kawaguchi Y
Department of Tumor Virology, Division of Virology and Immunology, Medical Research Institute, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8510, Japan.
Nakajima K
Igarashi M
Morita T
Tanaka M
Suzuki M
Yokoyama A
Matsuda G
Kato K
Kanamori M
Hirai K
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.21.10104-10111.2000
Published
2000-11-00
Pages
10104-11
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC102049
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.74.21.10104-10111.2000 PMC Full Text View journal details
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