Molecular cloning and characterization of an S-adenosylmethionine synthetase gene from Chorispora bungeana

Gene. 2015 Nov 10;572(2):205-13. doi: 10.1016/j.gene.2015.07.062. Epub 2015 Jul 20.

Abstract

S-adenosylmethionine synthetase (SAMS) catalyzes the formation of S-adenosylmethionine (SAM) which is a molecule essential for polyamines and ethylene biosynthesis, methylation modifications of protein, DNA and lipids. SAMS also plays an important role in abiotic stress response. Chorispora bungeana (C. bungeana) is an alpine subnival plant species which possesses strong tolerance to cold stress. Here, we cloned and characterized an S-adenosylmethionine synthetase gene, CbSAMS (C. bungeana S-adenosylmethionine synthetase), from C. bungeana, which encodes a protein of 393 amino acids containing a methionine binding motif GHPDK, an ATP binding motif GAGDQG and a phosphate binding motif GGGAFSGDK. Furthermore, an NES (nuclear export signal) peptide was identified through bioinformatics analysis. To explore the CbSAMS gene expression regulation, we isolated the promoter region of CbSAMS gene 1919bp upstream the ATG start codon, CbSAMSp, and analyzed its cis-acting elements by bioinformatics method. It was revealed that a transcription start site located at 320 bp upstream the ATG start codon and cis-acting elements related to light, ABA, auxin, ethylene, MeJA, low temperature and drought had been found in the CbSAMSp sequence. The gene expression pattern of CbSAMS was then analyzed by TR-qPCR and GUS assay method. The result showed that CbSAMS is expressed in all examined tissues including callus, roots, petioles, leaves, and flowers with a significant higher expression level in roots and flowers. Furthermore, the expression level of CbSAMS was induced by low temperature, ethylene and NaCl. Subcellular localization revealed that CbSAMS was located in the cytoplasm and nucleus but has a significant higher level in the nucleus. These results indicated a potential role of CbSAMS in abiotic stresses and plant growth in C. bungeana.

Keywords: Chorispora bungeana; Low temperature response; S-adenosylmethionine synthetase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Binding Sites
  • Brassicaceae / enzymology*
  • Brassicaceae / genetics
  • Cloning, Molecular / methods*
  • Computational Biology / methods
  • Flowers / genetics
  • Flowers / metabolism
  • Methionine Adenosyltransferase / chemistry
  • Methionine Adenosyltransferase / genetics*
  • Methionine Adenosyltransferase / metabolism*
  • Nuclear Export Signals / genetics
  • Plant Proteins / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plant Roots / genetics
  • Plant Roots / metabolism
  • Promoter Regions, Genetic

Substances

  • Nuclear Export Signals
  • Plant Proteins
  • Adenosine Triphosphate
  • Methionine Adenosyltransferase