ONE CARBON METABOLISM, THE METHIONINE CYCLE, AND HISTONE METHYLATION

Recent work has described a mechanism whereby the regulation of methionine metabolism affects methylation status in cells.  This regulation occurs through S-adenosylmethionine and its ability to potentiate methylation rreactions.  We have recently shown that Histone methylation is dramatically affected by changes in flux in the methionine cycle.  We are further investigating this phenomena.   

MASS SPECTROMETRY FOR QUANTITATIVE METABOLIC SIGNALING FLUX

We are using quantitative metabolomics approaches to measure to measure the methionine cycle flux under conditions of altered nutrient status.  These perturbations include differences in vitamins, amino acids, and glucose in controlled settings.  For each of these experiments stable isotope tracers will be considered and fluxes through methylation metabolism will be measured.  In addition the levels of metabolites in the methionine cycle will also be measured.    

DIRECT MEASUREMENTS OF SIGNALING FLUX FROM METABOLISM TO THE CHROMATIN

We will then connect the measured metabolite concentrations to signaling flux.  We will first measurement using quantitative western blotting of key modifications.  We will match the conditions under which we considered quantitative metabolomics and consider the state of histone modifications in these conditions.  To confirm the connections, we will measure and quantify the kinetics of signaling flux using isotopically labeled substrates.  Ultimately we will investigate the cancer context of this understudied form of cellular regulation.