The herpes simplex virus type 1 (HSV-1) UL35 open reading frame (ORF) encodes a 12-kDa capsid protein designated VP26. light, and therefore it is useful for analysis using fluorescence microscopy and circulation cytometry (5). Furthermore, this variant contains several silent base changes that correspond to optimal human codon usage for better expression in eukaryotic systems. This altered version of GFP (Clontech) was used for the experiments explained below. The aim of the experiments explained below was to incorporate the GFP into the HSV-1 capsid. A tagged nucleocapsid structure should be useful for the investigation of the Rabbit Polyclonal to ACRO (H chain, Cleaved-Ile43) early events in the uncoating of the computer virus particle and for monitoring the computer virus nucleocapsid during replication and transport in cell culture and in vivo. The rationale behind this approach was to utilize the VP26 polypeptide, which is located around the outer surface of the capsid shell. A fusion between the VP26 and GFP polypeptides was generated, and it was hoped that this fusion form of VP26 would still be capable of conversation with VP5 and would incorporate the GFP polypeptide onto the capsid structure. Consequently, the nucleocapsid and consequently the adult virion would be tagged with a fluorescent marker that is activated by light. Construction of a VP26-GFP fusion protein. The goal of the molecular manipulations explained below was to fuse the GFP ORF with that of VP26. An marker for viral replication. J Virol Methods. 1997;66:283C292. [PubMed] 14. McGeoch D J, Dalrymple M A, Davison A J, Dolan A, Frame M C, McNab D, Perry L J, Scott J E, Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1. J Gen Virol. 1988;69:1531C1574. [PubMed] 15. McNabb D S, Courtney R J. Identification and characterization of the herpes simplex virus type buy 1303607-60-4 1 virion protein encoded by the UL35 open reading frame. J Virol. 1992;66:2653C2663. [PMC free article] [PubMed] 16. McNabb D S, Courtney R J. Posttranslational modification and subcellular localization of the p12 capsid protein of herpes simplex buy 1303607-60-4 virus type 1. J Virol. 1992;66:4839C4847. [PMC free article] [PubMed] 17. Person S, Desai P. Capsids are created in a mutant computer virus blocked at the maturation site of the UL26 and UL26.5 open reading frames of HSV-1 but are not formed in a null mutant of UL38 (VP19C) Virology. 1998;242:193C203. [PubMed] 18. Rixon F J, Addison C, McGregor A, McNab S J, Nicholson P, Preston V G, Tatman J D. Multiple interactions control the intracellular localization of the herpes simplex virus type 1 capsid proteins. J Gen Virol. 1996;77:2251C2260. [PubMed] 19. Roizman B, Sears A. Herpes simplex viruses and their replication. In: Fields B N, Knipe D M, Howley P buy 1303607-60-4 M, et al., editors. Virology. Philadelphia, Pa: Lippincott-Raven; 1996. pp. 2231C2295. 20. Sodeik B, Ebersold M W, Helenius A. Microtubule-mediated transport of incoming herpes simplex virus type 1 capsids to the nucleus. J Cell Biol. 1997;136:1007C1021. [PMC free article] [PubMed] 21. Steven A C, Spear P G. Herpesvirus capsid assembly and envelopment. In: Burnett R, Chiu W, Garcea R, editors. Structural biology of viruses. New York, N.Y: Oxford University Press; 1996. pp. 312C351. 22. Tatman J D, Preston V G, Nicholson P, Elliot R M, Rixon F J. Assembly of herpes simplex virus type 1 capsids using a panel of recombinant baculoviruses. J Gen Virol. 1994;75:1101C1113. [PubMed] 23. Thomsen D R, Roof L L, Homa F L. Assembly of herpes simplex virus (HSV) intermediate capsids in insect cells infected with recombinant baculoviruses expressing HSV capsid proteins. J Virol. 1994;68:2442C2457. [PMC buy 1303607-60-4 free article] [PubMed] 24. Trus B L, Homa F L, Booy F P, Newcomb W W, Thomsen D R, Cheng N, Brown J C, Stevens A C. Herpes simplex virus capsids assembled in insect cells infected with recombinant baculoviruses: structural authenticity and localization of VP26. J Virol. 1995;69:7362C7366. buy 1303607-60-4 [PMC free article] [PubMed] 25. Ward P L, Ogle W O, Roizman B. Assemblons: nuclear structures defined by aggregation of immature capsids and some tegument proteins of herpes simplex virus type 1. J Virol. 1996;70:4623C4631. [PMC free article] [PubMed] 26. Wildy P, Russell W C, Horne R W. The morphology of herpes virus. Virology. 1960;12:204C222. [PubMed] 27. Wingfield P T, Stahl S J, Thomsen D R, Homa F L, Booy F P, Trus B L, Steven A C. Hexon-only binding of VP26 displays differences between the hexon and penton conformations of VP5, the major capsid protein of herpes simplex virus. J Virol. 1997;71:8955C8961. [PMC free article] [PubMed] 28. Zhou Z H, He J, Jakana J, Tatman J, Rixon F J, Chiu W. Assembly of VP26 in herpes simplex computer virus-1 inferred from structures of wild-type and recombinant capsids. Nat Struct Biol. 1995;2:1026C1030. [PubMed].