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Journal of Virology, May 2008, p. 5021-5030, Vol. 82, No. 10
0022-538X/08/$08.00+0     doi:10.1128/JVI.00150-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Domain within Herpes Simplex Virus 1 Scaffold Proteins Required for Interaction with Portal Protein in Infected Cells and Incorporation of the Portal Vertex into Capsids

Department of Microbiology and Immunology, New York State College of Veterinary Medicine, Cornell University, Ithaca, New York 14850

Received 21 January 2008/ Accepted 3 March 2008

The portal vertex of herpesvirus capsids serves as the conduit through which DNA is inserted during the assembly process. In herpes simplex virus (HSV), the portal is composed of 12 copies of the U_L6 gene product, pU_L6. Previous results identified a domain in the major capsid scaffold protein, ICP35, required for interaction with pU_L6 and its incorporation into capsids formed in vitro (G. P. Singer et al., J. Virol. 74:6838-6848, 2005). In the current studies, pU_L6 and scaffold proteins were found to coimmunoprecipitate from lysates of both HSV-infected cells and mammalian cells expressing scaffold proteins and pU_L6. The coimmunoprecipitation of pU_L6 and scaffold proteins was precluded upon deletion of codons 143 to 151 within U_L26.5, encoding ICP35. While wild-type scaffold proteins colocalized with pU_L6 when transiently coexpressed as viewed by indirect immunofluorescence, deletion of U_L26.5 codons 143 to 151 precluded this colocalization. A recombinant herpes simplex virus, vJB11, was generated that lacked U_L26.5 codons 143 to 151. A virus derived from this mutant but bearing a restored U_L26.5 was also generated. vJB11 was unable to cleave or package viral DNA, whereas the restored virus packaged DNA normally. vJB11 produced ample numbers of B capsids in infected cells, but these lacked normal levels of pU_L6. The deletion in U_L26.5 also rendered pU_L6 resistant to detergent extraction from vJB11-infected cells. These data indicate that, as was observed in vitro, amino acids 143 to 151 of ICP35 are critical for (i) interaction between scaffold proteins and pU_L6 and (ii) incorporation of the HSV portal into capsids.

Published ahead of print on 12 March 2008.

This article has been cited by other articles:

• Huffman, J. B., Newcomb, W. W., Brown, J. C., Homa, F. L. (2008). Amino Acids 143 to 150 of the Herpes Simplex Virus Type 1 Scaffold Protein Are Required for the Formation of Portal-Containing Capsids. J. Virol. 82: 6778-6781 [Abstract] [Full Text]