Possible regulation of trehalose metabolism by methylation in Saccharomyces cerevisiae.
The current study was undertaken to correlate post-translational protein modification by methylation with the functionality of enzymes involved in trehalose metabolism in Saccharomyces cerevisiae. Trehalose is an economically important disaccharide providing protection against various kinds of stresses. It also acts as a source of cellular energy by storing glucose. Methyl group donor S-adenosyl L-methionine (AdoMet) and methylation inhibitor-oxidized adenosine (AdOx) were used for the methylation study. AdoMet delayed initial growth of the cells but the overall growth rate remained same suggesting its interference in G1 phase of the cell cycle. Metabolic-altered enzyme activities of acid trehalase (AT), neutral trehalase (NT), and trehalose-6-phosphate synthase (TPS) were observed when treated with AdOx and AdoMet separately. A positive effect of methylation was observed in TPS, hence, it was purified in three different conditions, using AdoMet, AdOx, and control. Differences in mobility of methylated, methylation-inhibited, and control TPS during acidic native gel electrophoresis confirmed the occurrence of induced methylation. Hydrolysis under alkaline pH conditions revealed that methylation of TPS was different than O-methylation. MALDI-TOF analysis of trypsin-digested samples of purified methylated, methylation-inhibited, and control TPS revealed that an increase of 18 Da mass in methylated peptides suggesting the introduction of methyl ester in TPS. Results of amino acid analysis corroborated the presence of methyl cysteine. The data presented here strongly suggests that trehalose production was enhanced due to methylation of TPS arising from carboxymethylation of cysteine residues.
Sengupta
Shinjinee
S
Biotechnology Division, Indian Institute of Chemical Biology, Kolkata, West Bengal, India.
Chaudhuri
Paramita
P
Lahiri
Sagar
S
Dutta
Trina
T
Banerjee
Shakri
S
Majhi
Ramdhan
R
Ghosh
Anil K
AK
D000241:Adenosine / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D008745:Methylation; D011499:Protein Processing, Post-Translational; D012436:S-Adenosylmethionine / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014199:Trehalose / Q000378:metabolism*
D029701:Saccharomyces cerevisiae Proteins; D012436:S-Adenosylmethionine; D014199:Trehalose; D000241:Adenosine