Abstract
Reovirus genome segment S4 codes for polypeptide sigma3, a major outer capsid component of virions and a double-stranded RNA (dsRNA)-binding protein implicated in viral cytopathogenesis. We have constructed a stable HeLa cell line (S4tTA) that produces functional sigma3 under tetracycline transactivator control. In the absence of tetracycline, S4tTA cells synthesized stable dsRNA-binding sigma3 that accumulated in the nucleus as well as in the cytoplasm. However, in induced S4tTA cells also expressing reovirus outer shell polypeptide mu1/mu1C, migration of sigma3 into the nucleus was blocked, probably as a result of formation of a complex with mu1/mu1C which was exclusively in the cytoplasm. Mutant analyses indicated a correlation between dsRNA-binding activity and nuclear entry of sigma3, suggesting an additional role(s) for this capsid protein in virus-cell interactions.
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- Ahmed R., Fields B. N. Role of the S4 gene in the establishment of persistent reovirus infection in L cells. Cell. 1982 Mar;28(3):605–612. doi: 10.1016/0092-8674(82)90215-x. [DOI] [PubMed] [Google Scholar]
- Bernard H. U., Krämmer G., Röwekamp W. G. Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture. Exp Cell Res. 1985 May;158(1):237–243. doi: 10.1016/0014-4827(85)90446-x. [DOI] [PubMed] [Google Scholar]
- Bycroft M., Grünert S., Murzin A. G., Proctor M., St Johnston D. NMR solution structure of a dsRNA binding domain from Drosophila staufen protein reveals homology to the N-terminal domain of ribosomal protein S5. EMBO J. 1995 Jul 17;14(14):3563–3571. doi: 10.1002/j.1460-2075.1995.tb07362.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang H. W., Uribe L. H., Jacobs B. L. Rescue of vaccinia virus lacking the E3L gene by mutants of E3L. J Virol. 1995 Oct;69(10):6605–6608. doi: 10.1128/jvi.69.10.6605-6608.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen H. J., Remmler J., Delaney J. C., Messner D. J., Lobel P. Mutational analysis of the cation-independent mannose 6-phosphate/insulin-like growth factor II receptor. A consensus casein kinase II site followed by 2 leucines near the carboxyl terminus is important for intracellular targeting of lysosomal enzymes. J Biol Chem. 1993 Oct 25;268(30):22338–22346. [PubMed] [Google Scholar]
- Denzler K. L., Jacobs B. L. Site-directed mutagenic analysis of reovirus sigma 3 protein binding to dsRNA. Virology. 1994 Oct;204(1):190–199. doi: 10.1006/viro.1994.1523. [DOI] [PubMed] [Google Scholar]
- Gerace L. Nuclear export signals and the fast track to the cytoplasm. Cell. 1995 Aug 11;82(3):341–344. doi: 10.1016/0092-8674(95)90420-4. [DOI] [PubMed] [Google Scholar]
- Giantini M., Shatkin A. J. Stimulation of chloramphenicol acetyltransferase mRNA translation by reovirus capsid polypeptide sigma 3 in cotransfected COS cells. J Virol. 1989 Jun;63(6):2415–2421. doi: 10.1128/jvi.63.6.2415-2421.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gossen M., Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547–5551. doi: 10.1073/pnas.89.12.5547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huismans H., Joklik W. K. Reovirus-coded polypeptides in infected cells: isolation of two native monomeric polypeptides with affinity for single-stranded and double-stranded RNA, respectively. Virology. 1976 Apr;70(2):411–424. doi: 10.1016/0042-6822(76)90282-8. [DOI] [PubMed] [Google Scholar]
- Imani F., Jacobs B. L. Inhibitory activity for the interferon-induced protein kinase is associated with the reovirus serotype 1 sigma 3 protein. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7887–7891. doi: 10.1073/pnas.85.21.7887. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kharrat A., Macias M. J., Gibson T. J., Nilges M., Pastore A. Structure of the dsRNA binding domain of E. coli RNase III. EMBO J. 1995 Jul 17;14(14):3572–3584. doi: 10.1002/j.1460-2075.1995.tb07363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lloyd R. M., Shatkin A. J. Translational stimulation by reovirus polypeptide sigma 3: substitution for VAI RNA and inhibition of phosphorylation of the alpha subunit of eukaryotic initiation factor 2. J Virol. 1992 Dec;66(12):6878–6884. doi: 10.1128/jvi.66.12.6878-6884.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mabrouk T., Lemay G. Mutations in a CCHC zinc-binding motif of the reovirus sigma 3 protein decrease its intracellular stability. J Virol. 1994 Aug;68(8):5287–5290. doi: 10.1128/jvi.68.8.5287-5290.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Malim M. H., Böhnlein S., Hauber J., Cullen B. R. Functional dissection of the HIV-1 Rev trans-activator--derivation of a trans-dominant repressor of Rev function. Cell. 1989 Jul 14;58(1):205–214. doi: 10.1016/0092-8674(89)90416-9. [DOI] [PubMed] [Google Scholar]
- Mathews M. B., Shenk T. Adenovirus virus-associated RNA and translation control. J Virol. 1991 Nov;65(11):5657–5662. doi: 10.1128/jvi.65.11.5657-5662.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McCormack S. J., Thomis D. C., Samuel C. E. Mechanism of interferon action: identification of a RNA binding domain within the N-terminal region of the human RNA-dependent P1/eIF-2 alpha protein kinase. Virology. 1992 May;188(1):47–56. doi: 10.1016/0042-6822(92)90733-6. [DOI] [PubMed] [Google Scholar]
- Miller J. E., Samuel C. E. Proteolytic cleavage of the reovirus sigma 3 protein results in enhanced double-stranded RNA-binding activity: identification of a repeated basic amino acid motif within the C-terminal binding region. J Virol. 1992 Sep;66(9):5347–5356. doi: 10.1128/jvi.66.9.5347-5356.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pan L., Beverley P. C., Isaacson P. G. Lactate dehydrogenase (LDH) isoenzymes and proliferative activity of lymphoid cells--an immunocytochemical study. Clin Exp Immunol. 1991 Nov;86(2):240–245. doi: 10.1111/j.1365-2249.1991.tb05803.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Park H., Davies M. V., Langland J. O., Chang H. W., Nam Y. S., Tartaglia J., Paoletti E., Jacobs B. L., Kaufman R. J., Venkatesan S. TAR RNA-binding protein is an inhibitor of the interferon-induced protein kinase PKR. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4713–4717. doi: 10.1073/pnas.91.11.4713. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schiff L. A., Nibert M. L., Co M. S., Brown E. G., Fields B. N. Distinct binding sites for zinc and double-stranded RNA in the reovirus outer capsid protein sigma 3. Mol Cell Biol. 1988 Jan;8(1):273–283. doi: 10.1128/mcb.8.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seliger L. S., Giantini M., Shatkin A. J. Translational effects and sequence comparisons of the three serotypes of the reovirus S4 gene. Virology. 1992 Mar;187(1):202–210. doi: 10.1016/0042-6822(92)90308-c. [DOI] [PubMed] [Google Scholar]
- Sharpe A. H., Chen L. B., Fields B. N. The interaction of mammalian reoviruses with the cytoskeleton of monkey kidney CV-1 cells. Virology. 1982 Jul 30;120(2):399–411. doi: 10.1016/0042-6822(82)90040-x. [DOI] [PubMed] [Google Scholar]
- Sharpe A. H., Fields B. N. Reovirus inhibition of cellular RNA and protein synthesis: role of the S4 gene. Virology. 1982 Oct 30;122(2):381–391. doi: 10.1016/0042-6822(82)90237-9. [DOI] [PubMed] [Google Scholar]
- Smith R. E., Zweerink H. J., Joklik W. K. Polypeptide components of virions, top component and cores of reovirus type 3. Virology. 1969 Dec;39(4):791–810. doi: 10.1016/0042-6822(69)90017-8. [DOI] [PubMed] [Google Scholar]
- St Johnston D., Brown N. H., Gall J. G., Jantsch M. A conserved double-stranded RNA-binding domain. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10979–10983. doi: 10.1073/pnas.89.22.10979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tillotson L., Shatkin A. J. Reovirus polypeptide sigma 3 and N-terminal myristoylation of polypeptide mu 1 are required for site-specific cleavage to mu 1C in transfected cells. J Virol. 1992 Apr;66(4):2180–2186. doi: 10.1128/jvi.66.4.2180-2186.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Virgin H. W., 4th, Mann M. A., Fields B. N., Tyler K. L. Monoclonal antibodies to reovirus reveal structure/function relationships between capsid proteins and genetics of susceptibility to antibody action. J Virol. 1991 Dec;65(12):6772–6781. doi: 10.1128/jvi.65.12.6772-6781.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yuwen H., Cox J. H., Yewdell J. W., Bennink J. R., Moss B. Nuclear localization of a double-stranded RNA-binding protein encoded by the vaccinia virus E3L gene. Virology. 1993 Aug;195(2):732–744. doi: 10.1006/viro.1993.1424. [DOI] [PubMed] [Google Scholar]