Herpes simplex virus DNA cleavage and packaging:: Association of multiple forms of UL15-encoded proteins with B capsids requires at least the UL6, UL17, and UL28 genes

被引:50
作者
Salmon, B [1 ]
Baines, JD [1 ]
机构
[1] Cornell Univ, Dept Microbiol & Immunol, Vet Educ Ctr C5169, Ithaca, NY 14853 USA
关键词
D O I
10.1128/JVI.72.4.3045-3050.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The U(L)15 gene of herpes simpler virus (HSV) is one of several genes required for the packaging of viral DNA into intranuclear B capsids to produce C capsids that become enveloped at the inner nuclear membrane. A rabbit antiserum directed against ULS-encoded protein recognized three proteins with apparent M(r)s of 79,000, 80,000, and 83,000 in highly purified B capsids. The 83,000-M-r protein aas detected in type C capsids and comigrated with the product of a U(L)15 cDNA transcribed and translated in vitro. The 83,000- and 80,000-M-r proteins were readily detected in purified virions. Inasmuch as (i) none of these proteins were detectable in capsids purified from cells infected with HSV-1(Delta U(L)15), a virus lacking an intact U(L)15 gene, and (ii) corresponding proteins in capsids purified from cells infected with a recombinant virus [HSV-1(R7244), containing a 20-codon tag at the 3' end of U(L)15] were decreased in electrophoretic mobility relative to the wild-type proteins, we conclude that the proteins with apparent M(r)s of 83,000, 80,000, and 79,000 are products of U(L)15 with identical C termini. The 79,000-, 80,000-, and 83,000-M-r proteins remained associated with B capsids in the presence of 0.5 M guanidine HCl and remained detectable in capsids treated with 2.0 M guanidine HCl and lacking proteins associated with the capsid core. These data, therefore, indicate that U(L)15-encoded proteins are integral components of B capsids. Only the 83,000-M-r protein was detected in B capsids purified from cells infected with viruses lacking the U(L)6, U(L)17, or U(L)28 genes, which are required for DNA cleavage and packaging, suggesting that capsid association of the 80,000- and 79,000-M-r proteins requires intact cleavage and packaging machinery. These data, therefore, indicate that capsid association of the 80,000- and 79,000-M-r U(L)15-encoded proteins reflects a previously unrecognized step in the DNA cleavage and packaging reaction.
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页码:3045 / 3050
页数:6
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共 37 条
[21]   STRUCTURE OF THE HERPES-SIMPLEX VIRUS CAPSID - MOLECULAR COMPOSITION OF THE PENTONS AND THE TRIPLEXES [J].
NEWCOMB, WW ;
TRUS, BL ;
BOOY, FP ;
STEVEN, AC ;
WALL, JS ;
BROWN, JC .
JOURNAL OF MOLECULAR BIOLOGY, 1993, 232 (02) :499-511
[22]   STRUCTURE OF THE HERPES-SIMPLEX VIRUS CAPSID - EFFECTS OF EXTRACTION WITH GUANIDINE-HYDROCHLORIDE AND PARTIAL RECONSTITUTION OF EXTRACTED CAPSIDS [J].
NEWCOMB, WW ;
BROWN, JC .
JOURNAL OF VIROLOGY, 1991, 65 (02) :613-620
[23]   Assembly of the herpes simplex virus capsid: Characterization of intermediates observed during cell-free capsid formation [J].
Newcomb, WW ;
Homa, FL ;
Thomsen, DR ;
Booy, FP ;
Trus, BL ;
Steven, AC ;
Spencer, JV ;
Brown, JC .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 263 (03) :432-446
[24]   THE PRODUCT OF THE UL6 GENE OF HERPES-SIMPLEX VIRUS TYPE-1 IS ASSOCIATED WITH VIRUS CAPSIDS [J].
PATEL, AH ;
MACLEAN, JB .
VIROLOGY, 1995, 206 (01) :465-478
[25]   Isolation and characterization of herpes simplex virus type 1 mutants defective in the UL6 gene [J].
Patel, AH ;
Rixon, FJ ;
Cunningham, C ;
Davison, AJ .
VIROLOGY, 1996, 217 (01) :111-123
[26]   STUDIES OF MOLECULAR ANATOMY OF L-M CELL STRAIN OF EQUINE HERPES-VIRUS TYPE 1 - PROTEINS OF NUCLEOCAPSID AND INTACT VIRION [J].
PERDUE, ML ;
KEMP, MC ;
RANDALL, CC ;
OCALLAGH.DJ .
VIROLOGY, 1974, 59 (01) :201-216
[27]   CHARACTERIZATION OF 3 SPECIES OF NUCLEOCAPSIDS OF EQUINE HERPESVIRUS TYPE-1 (EHV-1) [J].
PERDUE, ML ;
COHEN, JC ;
KEMP, MC ;
RANDALL, CC ;
OCALLAGHAN, DJ .
VIROLOGY, 1975, 64 (01) :187-204
[28]   CHARACTERIZATION OF A TEMPERATURE-SENSITIVE MUTANT OF THE UL15 OPEN READING FRAME OF HERPES-SIMPLEX VIRUS-1 [J].
POON, APW ;
ROIZMAN, B .
JOURNAL OF VIROLOGY, 1993, 67 (08) :4497-4503
[29]   EVIDENCE THAT A PHAGE-T4 DNA PACKAGING ENZYME IS A PROCESSED FORM OF THE MAJOR CAPSID GENE-PRODUCT [J].
RAO, VB ;
BLACK, LW .
CELL, 1985, 42 (03) :967-977
[30]  
SALMON B, IN PRESS J VIROL