Molecular characterization of a rab-related small GTP binding protein cDNA from rice (Oryza sativa L. IR-36)

Mol Cells. 1997 Apr 30;7(2):226-30.

Abstract

To study physiological roles of plant small GTP binding proteins, we isolated a cDNA clone (ORrab2) encoding the rab-related small GTP binding protein from rice (Oryza sativa L. IR-36) by using human cDNA rab2 as a probe. The deduced amino acid sequence of the ORrab2 gene shared all the conserved regions, important for GTPase/GTP binding activities, with those of other small GTP binding proteins. ORrab2 is a 1028 bp long cDNA, encodes a 23.2 kDa protein which shows 85.2% similarity on the amino acid sequence level to the Hrab2 protein, and was used as a probe. Through Southern and Northern blot analyses, we found that ORrab2 is a single copy gene and actively expressed at the stages of cell division and elongation. We investigated GTP binding abilities by a filter assay procedure. Deletion of a binding motif, GDTGVGKS, within an ORrab2 protein showed a significant decrease of GTP binding affinity, suggesting its important role in nucleotide binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • DNA, Plant / genetics*
  • Escherichia coli / genetics
  • GTP-Binding Proteins / genetics*
  • GTP-Binding Proteins / metabolism
  • Gene Expression
  • Genes, Plant
  • Guanosine Triphosphate / metabolism
  • Humans
  • Molecular Sequence Data
  • Oryza / genetics*
  • Oryza / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • rab2 GTP-Binding Protein

Substances

  • DNA, Complementary
  • DNA, Plant
  • Plant Proteins
  • Guanosine Triphosphate
  • GTP-Binding Proteins
  • rab2 GTP-Binding Protein