The putative leucine zipper of the UL6-encoded portal protein of herpes simplex virus 1 is necessary for interaction with pUL15 and pUL28 and their association with capsids.
Herpes simplex virus (HSV) type 1 capsids contain a single portal vertex that is composed of 12 copies of the U(L)6 gene product (pU(L)6), which forms a pore through which DNA is inserted during packaging. This unique vertex is also believed to comprise the site with which a molecular motor, termed the terminase, associates during the DNA packaging reaction. In HSV, the terminase likely comprises the U(L)15, U(L)28, and U(L)33 proteins (pU(L)15, pU(L)28, and pU(L)33, respectively). The current study was undertaken to identify portal domains required for interaction with the terminase. Both the amino and carboxyl termini, as well as amino acids 422 to 443 of pU(L)6 forming a putative leucine zipper motif, were critical for coimmunoprecipitation with pU(L)15 in the absence of other viral proteins. Amino acids 422 to 443 were also necessary for interaction with pU(L)28 in the absence of other viral proteins. By using an engineered recombinant virus, it was further determined that although amino acids 422 to 443 were dispensable for interaction with scaffold protein and incorporation of portal protein into capsids, they were necessary for coimmunoprecipitation of pU(L)6 and pU(L)15 from infected cell lysates, association of optimal levels of pU(L)15, pU(L)28, and pU(L)33 with capsids, and DNA cleavage and packaging. These data identify a portal protein domain critical for terminase association with the capsid and suggest that both the pU(L)15- and pU(L)28-bearing terminase subunits mediate docking of the terminase with the portal vertex.
Yang
Kui
K
Department of Microbiology and Immunology, New York State College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
Wills
Elizabeth
E
Baines
Joel D
JD
D000818:Animals; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D000939:Epitopes / Q000276:immunology; D017353:Gene Deletion; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016350:Leucine Zippers; D011817:Rabbits; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014779:Virus Replication
D000939:Epitopes; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus