Abstract
Expression of rabbit hemorrhagic disease virus (RHDV) cDNAs in vitro with rabbit reticulocyte lysates and in Escherichia coli have been used to study the proteolytic processing of RHDV polyprotein encoded by ORF1. An epitope tag was used for monitoring the gene products by a specific antibody. We have identified four gene products with molecular masses of 80, 43, 73, and 60 kDa, from the amino to the carboxy terminus of the polyprotein. The amino-terminal sequences of the 43- and 73-kDa products were determined and indicated that RHDV 3C proteinase cleaved Glu-Gly peptide bonds.
Full Text
The Full Text of this article is available as a PDF (226.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allaire M., Chernaia M. M., Malcolm B. A., James M. N. Picornaviral 3C cysteine proteinases have a fold similar to chymotrypsin-like serine proteinases. Nature. 1994 May 5;369(6475):72–76. doi: 10.1038/369072a0. [DOI] [PubMed] [Google Scholar]
- Bazan J. F., Fletterick R. J. Viral cysteine proteases are homologous to the trypsin-like family of serine proteases: structural and functional implications. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7872–7876. doi: 10.1073/pnas.85.21.7872. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blok V. C., Wardell J., Jolly C. A., Manoukian A., Robinson D. J., Edwards M. L., Mayo M. A. The nucleotide sequence of RNA-2 of raspberry ringspot nepovirus. J Gen Virol. 1992 Sep;73(Pt 9):2189–2194. doi: 10.1099/0022-1317-73-9-2189. [DOI] [PubMed] [Google Scholar]
- Boga J. A., Casais R., Marin M. S., Martin-Alonso J. M., Carmenes R. S., Prieto M., Parra F. Molecular cloning, sequencing and expression in Escherichia coli of the capsid protein gene from rabbit haemorrhagic disease virus (Spanish isolate AST/89). J Gen Virol. 1994 Sep;75(Pt 9):2409–2413. doi: 10.1099/0022-1317-75-9-2409. [DOI] [PubMed] [Google Scholar]
- Boga J. A., Marín M. S., Casais R., Prieto M., Parra F. In vitro translation of a subgenomic mRNA from purified virions of the Spanish field isolate AST/89 of rabbit hemorrhagic disease virus (RHDV). Virus Res. 1992 Oct;26(1):33–40. doi: 10.1016/0168-1702(92)90144-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boniotti B., Wirblich C., Sibilia M., Meyers G., Thiel H. J., Rossi C. Identification and characterization of a 3C-like protease from rabbit hemorrhagic disease virus, a calicivirus. J Virol. 1994 Oct;68(10):6487–6495. doi: 10.1128/jvi.68.10.6487-6495.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
- Carter M. J., Milton I. D., Meanger J., Bennett M., Gaskell R. M., Turner P. C. The complete nucleotide sequence of a feline calicivirus. Virology. 1992 Sep;190(1):443–448. doi: 10.1016/0042-6822(92)91231-i. [DOI] [PubMed] [Google Scholar]
- Carter M. J., Milton I. D., Turner P. C., Meanger J., Bennett M., Gaskell R. M. Identification and sequence determination of the capsid protein gene of feline calicivirus. Arch Virol. 1992;122(3-4):223–235. doi: 10.1007/BF01317185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheah K. C., Leong L. E., Porter A. G. Site-directed mutagenesis suggests close functional relationship between a human rhinovirus 3C cysteine protease and cellular trypsin-like serine proteases. J Biol Chem. 1990 May 5;265(13):7180–7187. [PubMed] [Google Scholar]
- Forss S., Strebel K., Beck E., Schaller H. Nucleotide sequence and genome organization of foot-and-mouth disease virus. Nucleic Acids Res. 1984 Aug 24;12(16):6587–6601. doi: 10.1093/nar/12.16.6587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gauss-Müller V., Jürgensen D., Deutzmann R. Autoproteolytic cleavage of recombinant 3C proteinase of hepatitis A virus. Virology. 1991 Jun;182(2):861–864. doi: 10.1016/0042-6822(91)90630-t. [DOI] [PubMed] [Google Scholar]
- Gorbalenya A. E., Donchenko A. P., Blinov V. M., Koonin E. V. Cysteine proteases of positive strand RNA viruses and chymotrypsin-like serine proteases. A distinct protein superfamily with a common structural fold. FEBS Lett. 1989 Jan 30;243(2):103–114. doi: 10.1016/0014-5793(89)80109-7. [DOI] [PubMed] [Google Scholar]
- Harris K. S., Reddigari S. R., Nicklin M. J., Hämmerle T., Wimmer E. Purification and characterization of poliovirus polypeptide 3CD, a proteinase and a precursor for RNA polymerase. J Virol. 1992 Dec;66(12):7481–7489. doi: 10.1128/jvi.66.12.7481-7489.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jiang X., Wang M., Wang K., Estes M. K. Sequence and genomic organization of Norwalk virus. Virology. 1993 Jul;195(1):51–61. doi: 10.1006/viro.1993.1345. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Margis R., Pinck L. Effects of site-directed mutagenesis on the presumed catalytic triad and substrate-binding pocket of grapevine fanleaf nepovirus 24-kDa proteinase. Virology. 1992 Oct;190(2):884–888. doi: 10.1016/0042-6822(92)90931-e. [DOI] [PubMed] [Google Scholar]
- Margis R., Ritzenthaler C., Reinbolt J., Pinck M., Pinck L. Genome organization of grapevine fanleaf nepovirus RNA2 deduced from the 122K polyprotein P2 in vitro cleavage products. J Gen Virol. 1993 Sep;74(Pt 9):1919–1926. doi: 10.1099/0022-1317-74-9-1919. [DOI] [PubMed] [Google Scholar]
- Martín Alonso J. M., Balbín M., Garwes D. J., Enjuanes L., Gascón S., Parra F. Antigenic structure of transmissible gastroenteritis virus nucleoprotein. Virology. 1992 May;188(1):168–174. doi: 10.1016/0042-6822(92)90746-C. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
- Meyers G., Wirblich C., Thiel H. J. Genomic and subgenomic RNAs of rabbit hemorrhagic disease virus are both protein-linked and packaged into particles. Virology. 1991 Oct;184(2):677–686. doi: 10.1016/0042-6822(91)90437-G. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyers G., Wirblich C., Thiel H. J. Rabbit hemorrhagic disease virus--molecular cloning and nucleotide sequencing of a calicivirus genome. Virology. 1991 Oct;184(2):664–676. doi: 10.1016/0042-6822(91)90436-f. [DOI] [PubMed] [Google Scholar]
- Neill J. D. Nucleotide sequence of a region of the feline calicivirus genome which encodes picornavirus-like RNA-dependent RNA polymerase, cysteine protease and 2C polypeptides. Virus Res. 1990 Nov;17(3):145–160. doi: 10.1016/0168-1702(90)90061-f. [DOI] [PubMed] [Google Scholar]
- Ohlinger V. F., Haas B., Meyers G., Weiland F., Thiel H. J. Identification and characterization of the virus causing rabbit hemorrhagic disease. J Virol. 1990 Jul;64(7):3331–3336. doi: 10.1128/jvi.64.7.3331-3336.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palmenberg A. C. Proteolytic processing of picornaviral polyprotein. Annu Rev Microbiol. 1990;44:603–623. doi: 10.1146/annurev.mi.44.100190.003131. [DOI] [PubMed] [Google Scholar]
- Parra F., Boga J. A., Marin M. S., Casais R. The amino terminal sequence of VP60 from rabbit hemorrhagic disease virus supports its putative subgenomic origin. Virus Res. 1993 Mar;27(3):219–228. doi: 10.1016/0168-1702(93)90034-k. [DOI] [PubMed] [Google Scholar]
- Parra F., Prieto M. Purification and characterization of a calicivirus as the causative agent of a lethal hemorrhagic disease in rabbits. J Virol. 1990 Aug;64(8):4013–4015. doi: 10.1128/jvi.64.8.4013-4015.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
- Toyoda H., Nicklin M. J., Murray M. G., Anderson C. W., Dunn J. J., Studier F. W., Wimmer E. A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein. Cell. 1986 Jun 6;45(5):761–770. doi: 10.1016/0092-8674(86)90790-7. [DOI] [PubMed] [Google Scholar]
- Willcocks M. M., Brown T. D., Madeley C. R., Carter M. J. The complete sequence of a human astrovirus. J Gen Virol. 1994 Jul;75(Pt 7):1785–1788. doi: 10.1099/0022-1317-75-7-1785. [DOI] [PubMed] [Google Scholar]