en
Scientific article
English

Point Mutations in the Proximal Cys-His Box of Rous Sarcoma Virus Nucleocapsid Protein

Published inJournal of Virology, vol. 64, no. 10, p. 4978-4987
Publication date1990
Abstract

To extend our previous studies of the function of the Cys-His box of Rous sarcoma virus NC protein, we have constructed a series of point mutations of the conserved or nonconserved amino acids of the proximal Cys-His box and a one-amino-acid deletion. All mutants were characterized for production of viral proteins and particles, for packaging and maturation of viral RNA, for reverse transcriptase activity, and for infectivity. Our results indicated the following. (i) Mutations affecting the strictly conserved amino acids cysteine 21, cysteine 24, and histidine 29 were lethal; only the mutant His-29→Pro was still able to package viral RNA, most of it in an immature form. (ii) Mutation of the highly conserved glycine 28 to valine reduced viral RNA packaging by 90% and infectivity 30-fold, whereas mutant Gly-28→Ala was fully infectious. This suggests a steric hindrance limit at this position. (iii) Shortening the distance between cysteine 24 and histidine 29 by deleting one amino acid abolished the maturation of viral RNA and yielded noninfectious particles. (iv) Substitution of tyrosine 22 by serine lowered viral RNA packaging efficiency and yielded particles that were 400-fold less infectious; double mutant Tyr-22Thr-23→SerSer had the same infectivity as Tyr-22→Ser, whereas mutant Thr-23→Ser was fully infectious. (v) Changing glutamine 33 to a charged glutamate residue did not affect virus infectivity. Similarities and differences between our avian mutants and those in murine retroviruses are discussed.

Keywords
  • Amino Acid Sequence
  • Animals
  • Avian Sarcoma Viruses/genetics
  • Capsid/genetics
  • Cells
  • Cultured
  • Chick Embryo
  • Cysteine
  • Gene Products gag/genetics
  • Viral genes
  • Histidine
  • Molecular Sequence Data
  • Mutation
  • Oligonucleotide Probes
  • Viral RNA/genetics/isolation & purification
  • Transfection
  • Viral Core Proteins/genetics
  • Viral Proteins/genetics/isolation & purification
  • Virion/genetics
Funding
  • Swiss National Science Foundation - 3.066.087
Citation (ISO format)
DUPRAZ, Philippe et al. Point Mutations in the Proximal Cys-His Box of Rous Sarcoma Virus Nucleocapsid Protein. In: Journal of Virology, 1990, vol. 64, n° 10, p. 4978–4987. doi: 10.1128/JVI.64.10.4978-4987.1990
Main files (1)
Article (Published version)
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Identifiers
ISSN of the journal0022-538X
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