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A genetic screen for suppression of the growth defect of <jats:italic>Escherichia coli tktA tktB<\/jats:italic> mutant in LB medium revealed two mutations, one that rendered the <jats:italic>glpK<\/jats:italic> expression constitutive and another that inactivated <jats:italic>deoB<\/jats:italic>. Characterizing these mutations aided in uncovering the role of ribose\u20105\u2010P (a transketolase substrate) as an inhibitor of glycerol assimilation and <jats:italic>de novo<\/jats:italic> glycerol\u20103\u2010P synthesis. Using <jats:italic>lacZ<\/jats:italic> fusions, we show that ribose\u20105\u2010P enhances GlpR\u2013mediated repression of the <jats:italic>glpFKX<\/jats:italic> operon and inhibits glycerol assimilation. Electrophoretic Mobility Shift Assay (EMSA) showed ribose\u20105\u2010P made the DNA\u2010GlpR complex less sensitive to the inducer glycerol\u20103\u2010P. In addition to inhibition of glycerol assimilation, obstruction of ribose\u20105\u2010P metabolism retards growth from glycerol\u20103\u2010P limitation. Glucose helps to overcome this limitation through a mechanism involving catabolite repression. To our knowledge, this report is the first to show ribose\u20105\u2010P can modulate glycerol\u20103\u2010P concentration in the cell by regulation of glycerol assimilation as well as its <jats:italic>de novo<\/jats:italic> synthesis. This regulation could be prevalent in other organisms.<\/jats:p>","DOI":"10.1111\/mmi.13317","type":"journal-article","created":{"date-parts":[[2015,12,21]],"date-time":"2015-12-21T23:52:56Z","timestamp":1450741976000},"page":"263-277","source":"Crossref","is-referenced-by-count":21,"title":["Transketolase activity modulates glycerol\u20103\u2010phosphate levels in <scp><i>E<\/i><\/scp><i>scherichia coli<\/i>"],"prefix":"10.1111","volume":"100","author":[{"given":"A.","family":"Vimala","sequence":"first","affiliation":[{"name":"Laboratory of Bacterial Genetics Center for DNA Fingerprinting and Diagnostics Hyderabad 500 001 India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"Harinarayanan","sequence":"additional","affiliation":[{"name":"Laboratory of Bacterial Genetics Center for DNA Fingerprinting and Diagnostics Hyderabad 500 001 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