Cloning of the guanylate kinase homologues AGK-1 and AGK-2 from Arabidopsis thaliana and characterization of AGK-1

Eur J Biochem. 2000 Jan;267(2):606-15. doi: 10.1046/j.1432-1327.2000.01045.x.

Abstract

Guanylate kinase is an essential enzyme for nucleotide metabolism, phosphorylating GMP to GDP or dGMP to dGDP. The low molecular mass cytosolic forms of guanylate kinase are implicated primarily in the regulation of the supply of guanine nucleotides to cell signalling pathways. The high molecular mass and membrane-associated forms of guanylate kinase homologues, notably found in neuronal tissues, are assigned roles in cell junction organization and transmembrane regulation. Here, we describe the first plant guanylate kinase-encoding genes, AGK1 and AGK2, from Arabidopsis thaliana. The nucleotide sequences of their genomic and cDNA clones predict proteins that carry N-terminal and C-terminal extensions of the guanylate kinase-like domain. The amino acid sequences of this domain share 46-52% identity with guanylate kinases from yeast, Escherichia coli, human, mouse and Caenorhabditis elegans. Arabidopsis guanylate kinases (AGKs) exhibit a high degree of conservation of active site residues and sequence motifs in common with other nucleoside monophosphate kinases, which suggests overall structural similarity of the plant proteins. Although bacterially expressed AGK-1 is enzymatically much less active than yeast guanylate kinase, its kinase domain is shown to complement yeast GUK1 recessive lethal mutations. AGKs are expressed ubiquitously in plant tissues with highest transcriptional activity detected in roots. The identification of AGKs provides new perspectives for understanding the role of guanylate kinases in plant cell signalling pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arabidopsis / enzymology*
  • Arabidopsis / genetics
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Evolution, Molecular
  • Gene Dosage
  • Gene Expression Regulation, Plant
  • Genes, Fungal
  • Genetic Complementation Test
  • Guanylate Kinases
  • Humans
  • Mice
  • Molecular Sequence Data
  • Nucleoside-Phosphate Kinase / genetics*
  • Nucleoside-Phosphate Kinase / metabolism*
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism*
  • Plant Structures / enzymology
  • Plant Structures / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Yeasts / genetics

Substances

  • Plant Proteins
  • Recombinant Proteins
  • Nucleoside-Phosphate Kinase
  • Guanylate Kinases