E4orf6 variants with separate abilities to augment adenovirus replication and direct nuclear localization of the E1B 55-kilodalton protein

被引:19
作者
Orlando, JS [1 ]
Ornelles, DA [1 ]
机构
[1] Wake Forest Univ, Sch Med, Dept Microbiol & Immunol, Winston Salem, NC 27157 USA
关键词
D O I
10.1128/JVI.76.3.1475-1487.2002
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The E4orf6 protein of group C adenovirus is an oncoprotein that, in association with the E1B 55-kDa protein and by E1B-independent means, promotes virus replication. An arginine-faced amphipathic alpha-helix in the E4orf6 protein is required for the E4orf6 protein to direct nuclear localization of the E1B 55-kDa protein and to enhance replication of an E4 deletion virus. In this study, E4orf6 protein variants containing arginine substitutions in the amphipathic alpha-helix were analyzed. Two of the six arginine residues within the alpha-helix, arginine-241 and arginine-243, were critical for directing nuclear localization of the E1B 55-kDa protein. The four remaining arginine residues appear to provide a net positive charge for the E4orf6 protein to direct nuclear localization of the E1B 55-kDa protein. The molecular determinants of the arginine-faced amphipathic alpha-helix that were required for the functional interaction between the E4orf6 and E1B 55-kDa proteins seen in the transfected cell differed from those required to support a productive infection. Several E4orf6 protein variants with arginine-to-glutamic acid substitutions that failed to direct nuclear localization of the E1B 55-kDa protein restored replication of an E4 deletion virus. Additionally, a variant containing an arginine-to-alanine substitution at position 243 that directed nuclear localization of the E1B 55-kDa protein failed to enhance virus replication. These results indicate that the ability of the E4orf6 protein to relocalize the E1B 55-kDa protein to the nucleus can be separated from the ability of the E4orf6 protein to support a productive infection.
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页码:1475 / 1487
页数:13
相关论文
共 57 条
[1]   DNA-DAMAGING AGENTS GREATLY INCREASE THE TRANSDUCTION OF NONDIVIDING CELLS BY ADENOASSOCIATED VIRUS VECTORS [J].
ALEXANDER, IE ;
RUSSELL, DW ;
MILLER, AD .
JOURNAL OF VIROLOGY, 1994, 68 (12) :8282-8287
[2]   CHARACTERIZATION OF AN ADENOVIRUS GENE-TRANSFER VECTOR CONTAINING AN E4 DELETION [J].
ARMENTANO, D ;
SOOKDEO, CC ;
HEHIR, KM ;
GREGORY, RJ ;
STGEORGE, JA ;
PRINCE, GA ;
WADSWORTH, SC ;
SMITH, AE .
HUMAN GENE THERAPY, 1995, 6 (10) :1343-1353
[3]   SUPPRESSION OF HUMAN COLORECTAL-CARCINOMA CELL-GROWTH BY WILD-TYPE-P53 [J].
BAKER, SJ ;
MARKOWITZ, S ;
FEARON, ER ;
WILLSON, JKV ;
VOGELSTEIN, B .
SCIENCE, 1990, 249 (4971) :912-915
[4]  
Baldin V, 2000, Prog Cell Cycle Res, V4, P49
[5]   The N-terminus of Nef from HIV-1/SIV associates with a protein complex containing Lck and a serine kinase [J].
Baur, AS ;
Sass, G ;
Laffert, B ;
Willbold, D ;
ChengMayer, C ;
Peterlin, BM .
IMMUNITY, 1997, 6 (03) :283-291
[6]   Analysis of synthesis, stability, phosphorylation, and interacting polypeptides of the 34-kilodalton product of open reading frame 6 of the early region 4 protein of human adenovirus type 5 [J].
Boivin, D ;
Morrison, MR ;
Marcellus, RC ;
Querido, E ;
Branton, PE .
JOURNAL OF VIROLOGY, 1999, 73 (02) :1245-1253
[7]   A RAPID AND HIGHLY EFFICIENT METHOD FOR PCR-BASED SITE-DIRECTED MUTAGENESIS USING ONLY ONE NEW PRIMER [J].
BOLES, E ;
MIOSGA, T .
CURRENT GENETICS, 1995, 28 (02) :197-198
[8]   Adenovirus E4 34k and E4 11k inhibit double strand break repair and are physically associated with the cellular DNA-dependent protein kinase [J].
Boyer, J ;
Rohleder, K ;
Ketner, G .
VIROLOGY, 1999, 263 (02) :307-312
[9]   Genetic analysis of a potential zinc-binding domain of the adenovirus E4 34k protein [J].
Boyer, JL ;
Ketner, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (20) :14969-14978
[10]   INTERACTION OF ADENOVIRAL E4 AND E1B PRODUCTS IN LATE GENE-EXPRESSION [J].
BRIDGE, E ;
KETNER, G .
VIROLOGY, 1990, 174 (02) :345-353