Herpes simplex virus DNA cleavage and packaging: association of multiple forms of U(L)15-encoded proteins with B capsids requires at least the U(L)6, U(L)17, and U(L)28 genes.
The U(L)15 gene of herpes simplex 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 U(L)15-encoded protein recognized three proteins with apparent Mrs of 79,000, 80,000, and 83,000 in highly purified B capsids. The 83,000-Mr protein was 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-Mr 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(deltaU(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 Mrs 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-Mr 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-Mr 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-Mr proteins requires intact cleavage and packaging machinery. These data, therefore, indicate that capsid association of the 80,000- and 79,000-Mr U(L)15-encoded proteins reflects a previously unrecognized step in the DNA cleavage and packaging reaction.
Salmon
B
B
Department of Microbiology and Immunology, Cornell University, Ithaca, New York 14853, USA.
Baines
J D
JD
D000818:Animals; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D002522:Chlorocebus aethiops; D004279:DNA, Viral; D003902:Detergents / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D019791:Guanidine / Q000494:pharmacology; D018259:Herpesvirus 1, Human / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D006801:Humans; D016296:Mutagenesis; D011817:Rabbits; D014709:Vero Cells; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014771:Virion; D019065:Virus Assembly
D036022:Capsid Proteins; D004279:DNA, Viral; D003902:Detergents; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus; D019791:Guanidine