Proteomic analysis of a sea-ice diatom: salinity acclimation provides new insight into the dimethylsulfoniopropionate production pathway.
Dimethylsulfoniopropionate (DMSP) plays important roles in oceanic carbon and sulfur cycling and may significantly impact climate. It is a biomolecule synthesized from the methionine (Met) pathway and proposed to serve various physiological functions to aid in environmental stress adaptation through its compatible solute, cryoprotectant, and antioxidant properties. Yet, the enzymes and mechanisms regulating DMSP production are poorly understood. This study utilized a proteomics approach to investigate protein changes associated with salinity-induced DMSP increases in the model sea-ice diatom Fragilariopsis cylindrus (CCMP 1102). We hypothesized proteins associated with the Met-DMSP biosynthesis pathway would increase in relative abundance when challenged with elevated salinity. To test this hypothesis axenic log-phase cultures initially grown at a salinity of 35 were gradually shifted to a final salinity of 70 over a 24-h period. Intracellular DMSP was measured and two-dimensional gel electrophoresis was used to identify protein changes at 48 h after the shift. Intracellular DMSP increased by approximately 85% in the hypersaline cultures. One-third of the proteins increased under high salinity were associated with amino acid pathways. Three protein isoforms of S-adenosylhomo-cysteine hydrolase, which synthesizes a Met precursor, increased 1.8- to 2.1-fold, two isoforms of S-adenosyl Met synthetase increased 1.9- to 2.5-fold, and S-adenosyl Met methyltransferase increased by 2.8-fold, suggesting active methyl cycle proteins are recruited in the synthesis of DMSP. Proteins from the four enzyme classes of the proposed algal Met transaminase DMSP pathway were among the elevated proteins, supporting our hypothesis and providing candidate genes for future characterization studies.
Hollings Marine Lab, Charleston, South Carolina 29412.
Barbara R
BR
Lyon
Peter A
PA
Lee
Jennifer M
JM
Bennett
Giacomo R
GR
DiTullio
Michael G
MG
Janech
D000064:Acclimatization / Q000502:physiology*; D043383:Adenosylhomocysteinase / Q000378:metabolism; D017377:Diatoms / Q000201:enzymology / Q000254:growth & development / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D046448:Ice Cover; D008715:Methionine / Q000378:metabolism; D008716:Methionine Adenosyltransferase / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism; D010788:Photosynthesis; D045332:Photosystem II Protein Complex / Q000378:metabolism; D010839:Phytoplankton; D020033:Protein Isoforms; D011506:Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012436:S-Adenosylmethionine / Q000378:metabolism; D054712:Salinity; D013452:Sulfonium Compounds / Q000032:analysis / Q000378:metabolism*
D045332:Photosystem II Protein Complex; D020033:Protein Isoforms; D011506:Proteins; D013452:Sulfonium Compounds; D012436:S-Adenosylmethionine; D008715:Methionine; C068078:dimethylpropiothetin; D008780:Methyltransferases; D008716:Methionine Adenosyltransferase; D043383:Adenosylhomocysteinase