Herpes simplex virus DNA cleavage and packaging proteins associate with the procapsid prior to its maturation.
Packaging of DNA into preformed capsids is a fundamental early event in the assembly of herpes simplex virus type 1 (HSV-1) virions. Replicated viral DNA genomes, in the form of complex branched concatemers, and unstable spherical precursor capsids termed procapsids are thought to be the substrates for the DNA-packaging reaction. In addition, seven viral proteins are required for packaging, although their individual functions are undefined. By analogy to well-characterized bacteriophage systems, the association of these proteins with various forms of capsids, including procapsids, might be expected to clarify their roles in the packaging process. While the HSV-1 UL6, UL15, UL25, and UL28 packaging proteins are known to associate with different forms of stable capsids, their association with procapsids has not been tested. Therefore, we isolated HSV-1 procapsids from infected cells and used Western blotting to identify the packaging proteins present. Procapsids contained UL15 and UL28 proteins; the levels of both proteins are diminished in more mature DNA-containing C-capsids. In contrast, UL6 protein levels were approximately the same in procapsids, B-capsids, and C-capsids. The amount of UL25 protein was reduced in procapsids relative to that in more mature B-capsids. Moreover, C-capsids contained the highest level of UL25 protein, 15-fold higher than that in procapsids. Our results support current hypotheses on HSV DNA packaging: (i) transient association of UL15 and UL28 proteins with maturing capsids is consistent with their proposed involvement in site-specific cleavage of the viral DNA (terminase activity); (ii) the UL6 protein may be an integral component of the capsid shell; and (iii) the UL25 protein may associate with capsids after scaffold loss and DNA packaging, sealing the DNA within capsids.
Sheaffer
A K
AK
Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, Connecticut 06492, USA.
Newcomb
W W
WW
Gao
M
M
Yu
D
D
Weller
S K
SK
Brown
J C
JC
Tenney
D J
DJ
D000818:Animals; D015153:Blotting, Western; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D002460:Cell Line; D004279:DNA, Viral / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000235:genetics; D006561:Herpes Simplex / Q000821:virology; D009154:Mutation; D011498:Protein Precursors / Q000378:metabolism*; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism; D019065:Virus Assembly
D036022:Capsid Proteins; D004279:DNA, Viral; D011498:Protein Precursors; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus; D010450:Endopeptidases