This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sato, B. Articles by Arvin, A. M. Search for Related Content PubMed PubMed Citation Articles by Sato, B. Articles by Arvin, A. M.

Mutational Analysis of Open Reading Frames 62 and 71, Encoding the Varicella-Zoster Virus Immediate-Early Transactivating Protein, IE62, and Effects on Replication In Vitro and in Skin Xenografts in the SCID-hu Mouse In Vivo

Bunji Sato, Hideki Ito, Stewart Hinchliffe, Marvin H. Sommer, Leigh Zerboni, and Ann M. Arvin^*

Departments of Pediatrics and Microbiology, Stanford University School of Medicine, Stanford, California

Received 31 December 2002/ Accepted 13 February 2003

The varicella-zoster virus (VZV) genome has unique long (U_L) and unique short (U_S) segments which are flanked by internal repeat (IR) and terminal repeat (TR) sequences. The immediate-early 62 (IE62) protein, encoded by open reading frame 62 (ORF62) and ORF71 in these repeats, is the major VZV transactivating protein. Mutational analyses were done with VZV cosmids generated from parent Oka (pOka), a low-passage clinical isolate, and repair experiments were done with ORF62 from pOka and vaccine Oka (vOka), which is derived from pOka. Transfections using VZV cosmids from which ORF62, ORF71, or the ORF62/71 gene pair was deleted showed that VZV replication required at least one copy of ORF62. The insertion of ORF62 from pOka or vOka into a nonnative site in U_S allowed VZV replication in cell culture in vitro, although the plaque size and yields of infectious virus were decreased. Targeted mutations in binding sites reported to affect interaction with IE4 protein and a putative ORF9 protein binding site were not lethal. Single deletions of ORF62 or ORF71 from cosmids permitted recovery of infectious virus, but recombination events repaired the defective repeat region in some progeny viruses, as verified by PCR and Southern hybridization. VZV infectivity in skin xenografts in the SCID-hu model required ORF62 expression; mixtures of single-copy recombinant Oka62 (rOka62) or rOka71 and repaired rOka generated by recombination of the single-copy deletion mutants were detected in some skin implants. Although insertion of ORF62 into the nonnative site permitted replication in cell culture, ORF62 expression from its native site was necessary for cell-cell spread in differentiated human skin tissues in vivo.

Present address: Exploratory Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Tokodai, Tsukuba, Ibaraki 300-2698, Japan.

Present address: The London School of Tropical Medicine and Hygiene, London, United Kingdom.

This article has been cited by other articles:

• Zhang, Z., Rowe, J., Wang, W., Sommer, M., Arvin, A., Moffat, J., Zhu, H. (2007). Genetic Analysis of Varicella-Zoster Virus ORF0 to ORF4 by Use of a Novel Luciferase Bacterial Artificial Chromosome System. J. Virol. 81: 9024-9033 [Abstract] [Full Text]  
• Quinlivan, M. L., Gershon, A. A., Al Bassam, M. M., Steinberg, S. P., LaRussa, P., Nichols, R. A., Breuer, J. (2007). From the Cover: Natural selection for rash-forming genotypes of the varicella-zoster vaccine virus detected within immunized human hosts. Proc. Natl. Acad. Sci. USA 104: 208-212 [Abstract] [Full Text]  
• Hoover, S. E., Cohrs, R. J., Rangel, Z. G., Gilden, D. H., Munson, P., Cohen, J. I. (2006). Downregulation of Varicella-Zoster Virus (VZV) Immediate-Early ORF62 Transcription by VZV ORF63 Correlates with Virus Replication In Vitro and with Latency.. J. Virol. 80: 3459-3468 [Abstract] [Full Text]  
• Jones, J. O., Sommer, M., Stamatis, S., Arvin, A. M. (2006). Mutational Analysis of the Varicella-Zoster Virus ORF62/63 Intergenic Region. J. Virol. 80: 3116-3121 [Abstract] [Full Text]  
• Cohrs, R. J., Gilden, D. H., Gomi, Y., Yamanishi, K., Cohen, J. I. (2006). Comparison of Virus Transcription during Lytic Infection of the Oka Parental and Vaccine Strains of Varicella-Zoster Virus. J. Virol. 80: 2076-2082 [Abstract] [Full Text]  
• Taylor, S. L., Moffat, J. F. (2005). Replication of Varicella-Zoster Virus in Human Skin Organ Culture. J. Virol. 79: 11501-11506 [Abstract] [Full Text]  
• Ito, H., Sommer, M. H., Zerboni, L., Baiker, A., Sato, B., Liang, R., Hay, J., Ruyechan, W., Arvin, A. M. (2005). Role of the Varicella-Zoster Virus Gene Product Encoded by Open Reading Frame 35 in Viral Replication In Vitro and in Differentiated Human Skin and T Cells In Vivo. J. Virol. 79: 4819-4827 [Abstract] [Full Text]  
• Ku, C.-C., Besser, J., Abendroth, A., Grose, C., Arvin, A. M. (2005). Varicella-Zoster Virus Pathogenesis and Immunobiology: New Concepts Emerging from Investigations with the SCIDhu Mouse Model. J. Virol. 79: 2651-2658 [Full Text]  
• Besser, J., Ikoma, M., Fabel, K., Sommer, M. H., Zerboni, L., Grose, C., Arvin, A. M. (2004). Differential Requirement for Cell Fusion and Virion Formation in the Pathogenesis of Varicella-Zoster Virus Infection in Skin and T Cells. J. Virol. 78: 13293-13305 [Abstract] [Full Text]  
• Moffat, J., Mo, C., Cheng, J. J., Sommer, M., Zerboni, L., Stamatis, S., Arvin, A. M. (2004). Functions of the C-Terminal Domain of Varicella-Zoster Virus Glycoprotein E in Viral Replication In Vitro and Skin and T-Cell Tropism In Vivo. J. Virol. 78: 12406-12415 [Abstract] [Full Text]  
• Baiker, A., Fabel, K., Cozzio, A., Zerboni, L., Fabel, K., Sommer, M., Uchida, N., He, D., Weissman, I., Arvin, A. M. (2004). Varicella-zoster virus infection of human neural cells in vivo. Proc. Natl. Acad. Sci. USA 101: 10792-10797 [Abstract] [Full Text]  
• Baiker, A., Bagowski, C., Ito, H., Sommer, M., Zerboni, L., Fabel, K., Hay, J., Ruyechan, W., Arvin, A. M. (2004). The Immediate-Early 63 Protein of Varicella-Zoster Virus: Analysis of Functional Domains Required for Replication In Vitro and for T-Cell and Skin Tropism in the SCIDhu Model In Vivo. J. Virol. 78: 1181-1194 [Abstract] [Full Text]