Structure and assembly of turnip crinkle virus. VI. Identification of coat protein binding sites on the RNA

N. Wei, L. A. Heaton, Thomas J Morris, S. C. Harrison

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Structural studies of turnip crinkle virus have been extended to include the identification of high-affinity coat protein binding sites on the RNA genome. Virus was dissociated at elevated pH and ionic strength, and a ribonucleoprotein complex (rp-complex) was isolated by chromatography on Sephracyl S-200. Genomic RNA fragments in the rp-complex, resistant to RNase A and RNase T1 digestion and associated with tightly bound coat protein subunits, were isolated using coat-protein-specific antibodies. The identity of the protected fragments was determined by direct RNA sequencing. These approaches allowed us to study the specific RNA-protein interactions in the rp-complex obtained from dissociated virus particles. The location of one protected fragment downstream from the amber terminator codon in the first and largest of the three viral open reading frames suggests that the coat protein may play a role in the regulation of the expression of the polymerase gene. We have also identified an additional cluster of T1-protected fragments in the region of the coat protein gene that may represent further high-affinity sites involved in assembly recognition.

Original languageEnglish (US)
Pages (from-to)85-95
Number of pages11
JournalJournal of Molecular Biology
Volume214
Issue number1
DOIs
StatePublished - Jul 5 1990

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Carmovirus
Capsid Proteins
Protein Binding
Binding Sites
Ribonucleoproteins
RNA
Ribonuclease T1
RNA Sequence Analysis
Pancreatic Ribonuclease
Terminator Codon
Protein Subunits
Gene Expression Regulation
Virion
Osmolar Concentration
Open Reading Frames
Chromatography
Digestion
Genome
Viruses
Antibodies

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Structure and assembly of turnip crinkle virus. VI. Identification of coat protein binding sites on the RNA. / Wei, N.; Heaton, L. A.; Morris, Thomas J; Harrison, S. C.

In: Journal of Molecular Biology, Vol. 214, No. 1, 05.07.1990, p. 85-95.

Research output: Contribution to journalArticle

Wei, N. ; Heaton, L. A. ; Morris, Thomas J ; Harrison, S. C. / Structure and assembly of turnip crinkle virus. VI. Identification of coat protein binding sites on the RNA. In: Journal of Molecular Biology. 1990 ; Vol. 214, No. 1. pp. 85-95.
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