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PMID: 9223481 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The 222- to 234-kilodalton latent nuclear protein (LNA) of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) is encoded by orf73 and is a component of the latency-associated nuclear antigen.

Journal of virology ·Vol. 71 ·No. 8 ·1997-08-00 ·Pages 5915-21

Rainbow L, Platt GM, Simpson GR, Sarid R, Gao SJ, Stoiber H, Herrington CS, Moore PS, Schulz TF

Abstract

Kaposi's sarcoma (KS)-associated herpesvirus or human herpesvirus 8 (KSHV/HHV8) is the likely cause of KS and primary effusion lymphomas or body cavity-based lymphomas (BCBLs). A latency-associated nuclear immunofluorescence antigen (LANA) (D. H. Kedes, E. Operskalski, M. Busch, R. Kohn, J. Flood, and D. Ganem, Nat. Med. 2:918-924, 1996; S. J. Gao, L. Kingsley, M. Li, W. Zheng, C. Parravicini, J. Ziegler, R. Newton, C. R. Rinaldo, A. Saah, J. Phair, R. Detels, Y. Chang, and P. S. Moore, Nat. Med. 2:925-928, 1996) and a 222- to 234-kDa nuclear protein (LNA) (S. J. Gao, L. Kingsley, D. R. Hoover, T. J. Spira, C. R. Rinaldo, A. Saah, J. Phair, R. Detels, P. Parry, Y. Chang, and P. S. Moore, N. Engl. J. Med. 335:233-241, 1996) have previously been described in BCBL cell lines by immunofluorescence and Western blotting techniques, respectively. To identify the viral gene(s) encoding this antigen(s) we screened a cDNA library from HBL-6 cells, a B-cell lymphoma cell line persistently infected with KSHV/HHV8, with KS patient sera. One set of positive clones contained the 3' end of orf73, as well as the complete orf72 and orfK13, and another set contained the 5' end of orf73. Comparison of cDNA sequences with the KSHV/HHV8 genomic sequence revealed a splice event, occurring upstream of orf73. Immunoaffinity purified antibodies to a recombinant carboxy-terminal fragment of the orf73-encoded protein showed the characteristic speckled nuclear immunofluorescence pattern of LANA and reacted with the 222- to 234-kDa LNA on Western blots. Expression of full-length orf73 in bacteria and COS7 cells reproduced the LNA banding pattern. Immunohistochemistry on cases of nodular KS revealed that orf73/LNA is expressed in the nucleus of KS spindle cells. These findings demonstrate that orf73 encodes the 222- to 234-kDa LNA, is a component of LANA, and is expressed in KS tumor cells.

MeSH Terms
Animals Antigens, Nuclear/genetics,physiology Blotting, Western COS Cells DNA, Complementary/analysis Fluorescent Antibody Technique Herpesvirus 8, Human/chemistry,genetics Molecular Weight Nuclear Proteins/genetics,physiology Open Reading Frames Sarcoma, Kaposi/virology Viral Proteins/genetics,physiology
Chemicals
Antigens, Nuclear DNA, Complementary Nuclear Proteins Viral Proteins latent Orf73 antigen, human herpesvirus 8
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Rainbow L
Department of Medical Microbiology and Genitourinary Medicine, The University of Liverpool, United Kingdom.
Platt G M
Simpson G R
Sarid R
Gao S J
Stoiber H
Herrington C S
Moore P S
Schulz T F
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.71.8.5915-5921.1997
Published
1997-08-00
Pages
5915-21
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC191847
Subset
IM
Grants
PHS HHS · U64CCU210852 · United States
External Links
PubMed source DOI: 10.1128/JVI.71.8.5915-5921.1997 PMC Full Text View journal details
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