Comparison of the nucleic acid- and NTP-binding properties of the movement protein of cucumber mosaic cucumovirus and tobacco mosaic tobamovirus

Virology. 1996 Feb 1;216(1):71-9. doi: 10.1006/viro.1996.0035.

Abstract

The cucumber mosaic virus (CMV) 3a movement protein (MP) was compared directly to the well-characterized tobacco mosaic virus (TMV) 30K MP by cloning the genes encoding these proteins into Escherichia coli, isolating the E. coli-expressed MPs, and characterizing them with regard to RNA- and NTP-binding activities. The two MPs were shown to bind single-stranded RNA and DNA cooperatively, but with no sequence specificity. However, discrete lengths of CMV RNA 3 could be protected against RNase digestion by the CMV 3a protein, indicating that the RNA was not uniformly covered by the MP after cooperative binding. The TMV 30K:RNA complex was more stable in NaCl than the CMV 3a:RNA complex; about 50% of the corresponding complexes were stable in 0.6 and 0.4 M NaCl, respectively. Both MPs could bind GTP strongly and UTP weakly, but not ATP or CTP. The CMV 3a protein expressed either in E. coli or in planta from RNA 3 of CMV was tagged at its C-terminus with six histidine residues, which facilitated its purification by affinity chromatography on a matrix containing Ni(2+)-nitrilotriacetate. The soluble, His-tagged 3a proteins, affinity-purified from E. coli and zucchini squash, both were able bind CMV RNA 3 in vitro.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • Cucumovirus / genetics
  • Cucumovirus / metabolism*
  • DNA, Viral / metabolism
  • DNA-Binding Proteins / metabolism
  • Escherichia coli
  • Histidine / metabolism
  • Molecular Sequence Data
  • Nucleotides / metabolism*
  • Plant Viral Movement Proteins
  • Plants / virology
  • Protein Binding
  • RNA, Viral / metabolism*
  • RNA-Binding Proteins / metabolism*
  • Recombinant Proteins / metabolism
  • Sodium Chloride / pharmacology
  • Tobacco Mosaic Virus / genetics
  • Tobacco Mosaic Virus / metabolism*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • DNA, Viral
  • DNA-Binding Proteins
  • Nucleotides
  • Plant Viral Movement Proteins
  • RNA, Viral
  • RNA-Binding Proteins
  • Recombinant Proteins
  • Viral Proteins
  • Sodium Chloride
  • Histidine