Abstract
Earlier studies have shown that the UL11 gene of herpes simplex virus encodes a myristylated virion protein and that the UL11 gene enables efficient virion envelopment and export from infected cells. A rabbit polyclonal antibody directed against an affinity-purified UL11-glutathione-S-transferase fusion protein was made and used to study the properties of the UL11 protein and its distribution in infected cells. We report the following: (i) UL11 protein formed up to five bands (apparent M(r)s, 17,000 to 22,000) in denaturing polyacrylamide gels; (ii) fluorescent-antibody studies revealed the presence of UL11 protein in the perinuclear space and in sites within the nucleus; (iii) immune electron microscopic studies indicated that the UL11 gene products were associated with the inner nuclear membrane, with cytoplasmic membranes and ribbon-like cytoplasmic structures resembling membranous organelles, with nuclear bodies shown by fluorescence microscopy to be different from nucleoli in which US11 protein accumulates, and with enveloped virions but not with nuclear capsids; and (iv) the nuclear bodies containing UL11 protein were reminiscent both of type IV morphotypes consisting of an electron-dense core containing the UL11 proteins surrounded by a more electron-transluscent core and of type V morphotypes consisting of material homogenous in electron opacity. We conclude that (i) the UL11 protein is processed after synthesis; (ii) the localization of UL11 protein with virions and membranes is consistent with the hypothesis that UL11 plays a role in the transport of virions to the extracellular space; and (iii) although the significance of the association of UL11 proteins with nuclear bodies is unknown, the results indicate that nuclear bodies differ with respect to their morphologies and contents of viral protein and suggest that UL11 protein may have more than one function in the infected cell.
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- Ackermann M., Chou J., Sarmiento M., Lerner R. A., Roizman B. Identification by antibody to a synthetic peptide of a protein specified by a diploid gene located in the terminal repeats of the L component of herpes simplex virus genome. J Virol. 1986 Jun;58(3):843–850. doi: 10.1128/jvi.58.3.843-850.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baines J. D., Roizman B. The UL10 gene of herpes simplex virus 1 encodes a novel viral glycoprotein, gM, which is present in the virion and in the plasma membrane of infected cells. J Virol. 1993 Mar;67(3):1441–1452. doi: 10.1128/jvi.67.3.1441-1452.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baines J. D., Roizman B. The UL11 gene of herpes simplex virus 1 encodes a function that facilitates nucleocapsid envelopment and egress from cells. J Virol. 1992 Aug;66(8):5168–5174. doi: 10.1128/jvi.66.8.5168-5174.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baines J. D., Ward P. L., Campadelli-Fiume G., Roizman B. The UL20 gene of herpes simplex virus 1 encodes a function necessary for viral egress. J Virol. 1991 Dec;65(12):6414–6424. doi: 10.1128/jvi.65.12.6414-6424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baradaran K., Dabrowski C. E., Schaffer P. A. Transcriptional analysis of the region of the herpes simplex virus type 1 genome containing the UL8, UL9, and UL10 genes and identification of a novel delayed-early gene product, OBPC. J Virol. 1994 Jul;68(7):4251–4261. doi: 10.1128/jvi.68.7.4251-4261.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barker D. E., Roizman B. The unique sequence of the herpes simplex virus 1 L component contains an additional translated open reading frame designated UL49.5. J Virol. 1992 Jan;66(1):562–566. doi: 10.1128/jvi.66.1.562-566.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bouteille M., Kalifat S. R., Delarue J. Ultrastructural variations of nuclear bodies in human diseases. J Ultrastruct Res. 1967 Aug 30;19(5):474–486. doi: 10.1016/s0022-5320(67)80074-1. [DOI] [PubMed] [Google Scholar]
- Brasch K., Ochs R. L. Nuclear bodies (NBs): a newly "rediscovered" organelle. Exp Cell Res. 1992 Oct;202(2):211–223. doi: 10.1016/0014-4827(92)90068-j. [DOI] [PubMed] [Google Scholar]
- Bush M., Yager D. R., Gao M., Weisshart K., Marcy A. I., Coen D. M., Knipe D. M. Correct intranuclear localization of herpes simplex virus DNA polymerase requires the viral ICP8 DNA-binding protein. J Virol. 1991 Mar;65(3):1082–1089. doi: 10.1128/jvi.65.3.1082-1089.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campadelli-Fiume G., Farabegoli F., Di Gaeta S., Roizman B. Origin of unenveloped capsids in the cytoplasm of cells infected with herpes simplex virus 1. J Virol. 1991 Mar;65(3):1589–1595. doi: 10.1128/jvi.65.3.1589-1595.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou J., Roizman B. The herpes simplex virus 1 gene for ICP34.5, which maps in inverted repeats, is conserved in several limited-passage isolates but not in strain 17syn+. J Virol. 1990 Mar;64(3):1014–1020. doi: 10.1128/jvi.64.3.1014-1020.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou J., Roizman B. The terminal a sequence of the herpes simplex virus genome contains the promoter of a gene located in the repeat sequences of the L component. J Virol. 1986 Feb;57(2):629–637. doi: 10.1128/jvi.57.2.629-637.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Desai P. J., Schaffer P. A., Minson A. C. Excretion of non-infectious virus particles lacking glycoprotein H by a temperature-sensitive mutant of herpes simplex virus type 1: evidence that gH is essential for virion infectivity. J Gen Virol. 1988 Jun;69(Pt 6):1147–1156. doi: 10.1099/0022-1317-69-6-1147. [DOI] [PubMed] [Google Scholar]
- Ejercito P. M., Kieff E. D., Roizman B. Characterization of herpes simplex virus strains differing in their effects on social behaviour of infected cells. J Gen Virol. 1968 May;2(3):357–364. doi: 10.1099/0022-1317-2-3-357. [DOI] [PubMed] [Google Scholar]
- Georgopoulou U., Michaelidou A., Roizman B., Mavromara-Nazos P. Identification of a new transcriptional unit that yields a gene product within the unique sequences of the short component of the herpes simplex virus 1 genome. J Virol. 1993 Jul;67(7):3961–3968. doi: 10.1128/jvi.67.7.3961-3968.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lagunoff M., Roizman B. Expression of a herpes simplex virus 1 open reading frame antisense to the gamma(1)34.5 gene and transcribed by an RNA 3' coterminal with the unspliced latency-associated transcript. J Virol. 1994 Sep;68(9):6021–6028. doi: 10.1128/jvi.68.9.6021-6028.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu F. Y., Roizman B. The promoter, transcriptional unit, and coding sequence of herpes simplex virus 1 family 35 proteins are contained within and in frame with the UL26 open reading frame. J Virol. 1991 Jan;65(1):206–212. doi: 10.1128/jvi.65.1.206-212.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacLean C. A., Clark B., McGeoch D. J. Gene UL11 of herpes simplex virus type 1 encodes a virion protein which is myristylated. J Gen Virol. 1989 Dec;70(Pt 12):3147–3157. doi: 10.1099/0022-1317-70-12-3147. [DOI] [PubMed] [Google Scholar]
- MacLean C. A., Dolan A., Jamieson F. E., McGeoch D. J. The myristylated virion proteins of herpes simplex virus type 1: investigation of their role in the virus life cycle. J Gen Virol. 1992 Mar;73(Pt 3):539–547. doi: 10.1099/0022-1317-73-3-539. [DOI] [PubMed] [Google Scholar]
- MacLean C. A., Rixon F. J., Marsden H. S. The products of gene US11 of herpes simplex virus type 1 are DNA-binding and localize to the nucleoli of infected cells. J Gen Virol. 1987 Jul;68(Pt 7):1921–1937. doi: 10.1099/0022-1317-68-7-1921. [DOI] [PubMed] [Google Scholar]
- Mackem S., Roizman B. Regulation of herpesvirus macromolecular synthesis: transcription-initiation sites and domains of alpha genes. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7122–7126. doi: 10.1073/pnas.77.12.7122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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 Jul;69(Pt 7):1531–1574. doi: 10.1099/0022-1317-69-7-1531. [DOI] [PubMed] [Google Scholar]
- McGeoch D. J., Dolan A., Donald S., Rixon F. J. Sequence determination and genetic content of the short unique region in the genome of herpes simplex virus type 1. J Mol Biol. 1985 Jan 5;181(1):1–13. doi: 10.1016/0022-2836(85)90320-1. [DOI] [PubMed] [Google Scholar]
- Post L. E., Roizman B. A generalized technique for deletion of specific genes in large genomes: alpha gene 22 of herpes simplex virus 1 is not essential for growth. Cell. 1981 Jul;25(1):227–232. doi: 10.1016/0092-8674(81)90247-6. [DOI] [PubMed] [Google Scholar]
- Puvion-Dutilleul F. Localization of viral-specific 21 kDa protein in nucleoli of herpes simplex infected cells. Eur J Cell Biol. 1987 Jun;43(3):487–498. [PubMed] [Google Scholar]
- Puvion-Dutilleul F., Pichard E. Viral alkaline nuclease in intranuclear dense bodies induced by herpes simplex infection. Biol Cell. 1986;58(1):15–22. doi: 10.1111/j.1768-322x.1986.tb00485.x. [DOI] [PubMed] [Google Scholar]
- Rixon F. J., Atkinson M. A., Hay J. Intranuclear distribution of herpes simplex virus type 2 DNA synthesis: examination by light and electron microscopy. J Gen Virol. 1983 Sep;64(Pt 9):2087–2092. doi: 10.1099/0022-1317-64-9-2087. [DOI] [PubMed] [Google Scholar]
- Roller R. J., Roizman B. The herpes simplex virus 1 RNA binding protein US11 is a virion component and associates with ribosomal 60S subunits. J Virol. 1992 Jun;66(6):3624–3632. doi: 10.1128/jvi.66.6.3624-3632.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz J., Roizman B. Similarities and Differences in the Development of Laboratory Strains and Freshly Isolated Strains of Herpes Simplex Virus in HEp-2 Cells: Electron Microscopy. J Virol. 1969 Dec;4(6):879–889. doi: 10.1128/jvi.4.6.879-889.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shao L., Rapp L. M., Weller S. K. Herpes simplex virus 1 alkaline nuclease is required for efficient egress of capsids from the nucleus. Virology. 1993 Sep;196(1):146–162. doi: 10.1006/viro.1993.1463. [DOI] [PubMed] [Google Scholar]
- Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
- de THE, RIVIERE M., BERNHARD W. [Examination by electron microscope of the VX2 tumor of the domestic rabbit derived from the Shope papilloma]. Bull Assoc Fr Etud Cancer. 1960 Oct-Dec;47:570–584. [PubMed] [Google Scholar]