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. 1982 May;42(2):630–635. doi: 10.1128/jvi.42.2.630-635.1982

Expression of the Bottom-Component RNA of Cowpea Mosaic Virus: Evidence that the 60-Kilodalton VPg Precursor Is Cleaved into Single VPg and a 58-Kilodalton Polypeptide

Rob Goldbach 1, Geertje Rezelman 1, Pim Zabel 1, Albert van Kammen 1
PMCID: PMC256888  PMID: 16789223

Abstract

In cowpea protoplasts infected with cowpea mosaic virus, a bottom-component (B) RNA-encoded 60-kilodalton (60K) polypeptide is synthesized, which is membrane-bound and represents the direct precursor to the genome-bound protein VPg. The relationship between this VPg precursor and other B-RNA-encoded polypeptides was studied. Digestion of the B-RNA-encoded 170K and 84K polypeptides with Staphylococcus aureus protease V8 and subsequent analysis of the generated peptides with antiserum against VPg showed that a VPg sequence resides internally in these polypeptides. Furthermore, a new B-RNA-encoded polypeptide was detected, with a size of 58K, which differed from the 60K polypeptide only in the lack of VPg sequences. A model is presented in which the 60K VPg precursor is generated from the 200K primary translation product from B RNA and further processed to a 58K polypeptide and single VPg.

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Selected References

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  1. Ahlquist P., Kaesberg P. Determination of the length distribution of poly(A) at the 3' terminus of the virion RNAs of EMC virus, poliovirus, rhinovirus, RAV-61 and CPMV and of mouse globin mRNA. Nucleic Acids Res. 1979 Nov 10;7(5):1195–1204. doi: 10.1093/nar/7.5.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Daubert S. D., Bruening G., Najarian R. C. Protein bound to the genome RNAs of cowpea mosaic virus. Eur J Biochem. 1978 Dec 1;92(1):45–51. doi: 10.1111/j.1432-1033.1978.tb12721.x. [DOI] [PubMed] [Google Scholar]
  3. Franssen H., Goldbach R., Broekhuijsen M., Moerman M., van Kammen A. Expression of Middle-Component RNA of Cowpea Mosaic Virus: In Vitro Generation of a Precursor to Both Capsid Proteins by a Bottom-Component RNA-Encoded Protease from Infected Cells. J Virol. 1982 Jan;41(1):8–17. doi: 10.1128/jvi.41.1.8-17.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hibi T., Rezelman G., Van Kammen A. Infection of cowpea mesophyll protoplasts with cowpea mosaic virus. Virology. 1975 Apr;64(2):308–318. doi: 10.1016/0042-6822(75)90107-5. [DOI] [PubMed] [Google Scholar]
  5. Kitamura N., Adler C. J., Rothberg P. G., Martinko J., Nathenson S. G., Wimmer E. The genome-linked protein of picornaviruses. VII. Genetic mapping of poliovirus VPg by protein and RNA sequence studies. Cell. 1980 Aug;21(1):295–302. doi: 10.1016/0092-8674(80)90137-3. [DOI] [PubMed] [Google Scholar]
  6. Klootwijk J., Klein I., Zabel P., van Kammen A. Cowpea mosaic virus RNAs have neither m7GpppN ... nor mono-, di- or triphosphates at their 5' ends. Cell. 1977 May;11(1):73–82. doi: 10.1016/0092-8674(77)90318-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Pallansch M. A., Kew O. M., Palmenberg A. C., Golini F., Wimmer E., Rueckert R. R. Picornaviral VPg sequences are contained in the replicase precursor. J Virol. 1980 Aug;35(2):414–419. doi: 10.1128/jvi.35.2.414-419.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pettersson R. F., Ambros V., Baltimore D. Identification of a protein linked to nascent poliovirus RNA and to the polyuridylic acid of negative-strand RNA. J Virol. 1978 Aug;27(2):357–365. doi: 10.1128/jvi.27.2.357-365.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rezelman G., Goldbach R., Van Kammen A. Expression of bottom component RNA of cowpea mosaic virus in cowpea protoplasts. J Virol. 1980 Nov;36(2):366–373. doi: 10.1128/jvi.36.2.366-373.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Stanley J., Rottier P., Davies J. W., Zabel P., Van Kammen A. A protein linked to the 5' termini of both RNA components of the cowpea mosaic virus genome. Nucleic Acids Res. 1978 Dec;5(12):4505–4522. doi: 10.1093/nar/5.12.4505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Zabel P., Moerman M., van Straaten F., Goldbach R., van Kammen A. Antibodies Against the Genome-Linked Protein VPg of Cowpea Mosaic Virus Recognize a 60,000-Dalton Precursor Polypeptide. J Virol. 1982 Mar;41(3):1083–1088. doi: 10.1128/jvi.41.3.1083-1088.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. el-Manna M. M., Bruening G. Polyadenylate sequences in the ribonucleic acids of cowpea mosaic virus. Virology. 1973 Nov;56(1):198–206. doi: 10.1016/0042-6822(73)90299-7. [DOI] [PubMed] [Google Scholar]
  14. van Kammen A. Purification and properties of the components of cowpea mosaic virus. Virology. 1967 Apr;31(4):633–642. doi: 10.1016/0042-6822(67)90192-4. [DOI] [PubMed] [Google Scholar]

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