Relationship between Growth Rate and ATP Concentration in Escherichia coli: A BIOASSAY FOR AVAILABLE CELLULAR ATP Export

@article{citeulike:2460926, abstract = {Previous studies showed that adenosine triphosphate (ATP) concentrations in Escherichia coli changed during certain growth transitions and directly controlled the rate of rRNA transcription initiation at those times. The relationship between ATP concentration and rRNA transcription during steady-state growth is less clear, however. This is because two commonly employed methods for measuring ATP concentrations in bacteria, both of which rely on physical extraction followed by chromatographic separation of small molecules, resulted in dramatically different conclusions about whether ATP concentration changed with steady-state growth rate. Extraction with formic acid indicated that ATP concentration did not change with growth rate, whereas formaldehyde treatment followed by extraction with alkali indicated that ATP concentration increased proportionally to the growth rate. To resolve this discrepancy, we developed a bioassay for ATP based on the expression of a variant of the firefly luciferase enzyme in vivo and measurement of luminescence in cells growing in different conditions. We found that the available ATP concentration did not vary with growth rate, either in wild-type cells or in cells lacking guanosine 5'-diphosphate, 3'-diphosphate, providing insight into the regulation of rRNA transcription. More broadly, the luciferase bioassay described here provides a general method for evaluating the ATP concentration available for biochemical processes in E. coli and potentially in other organisms. 10.1074/jbc.M311996200}, author = {Schneider, David A. and Gourse, Richard L.}, citeulike-article-id = {2460926}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M311996200}, citeulike-linkout-1 = {http://www.jbc.org/cgi/content/abstract/279/9/8262}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/14670952}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=14670952}, day = {27}, doi = {10.1074/jbc.M311996200}, journal = {J. Biol. Chem.}, keywords = {atp, ecoli, growth, sampling}, month = {February}, number = {9}, pages = {8262--8268}, posted-at = {2008-03-03 14:15:23}, priority = {2}, title = {Relationship between Growth Rate and ATP Concentration in Escherichia coli: A BIOASSAY FOR AVAILABLE CELLULAR ATP}, url = {http://dx.doi.org/10.1074/jbc.M311996200}, volume = {279}, year = {2004} }

TY - JOUR ID - citeulike:2460926 L3 - citeulike-article-id:2460926 N2 - Previous studies showed that adenosine triphosphate (ATP) concentrations in Escherichia coli changed during certain growth transitions and directly controlled the rate of rRNA transcription initiation at those times. The relationship between ATP concentration and rRNA transcription during steady-state growth is less clear, however. This is because two commonly employed methods for measuring ATP concentrations in bacteria, both of which rely on physical extraction followed by chromatographic separation of small molecules, resulted in dramatically different conclusions about whether ATP concentration changed with steady-state growth rate. Extraction with formic acid indicated that ATP concentration did not change with growth rate, whereas formaldehyde treatment followed by extraction with alkali indicated that ATP concentration increased proportionally to the growth rate. To resolve this discrepancy, we developed a bioassay for ATP based on the expression of a variant of the firefly luciferase enzyme in vivo and measurement of luminescence in cells growing in different conditions. We found that the available ATP concentration did not vary with growth rate, either in wild-type cells or in cells lacking guanosine 5'-diphosphate, 3'-diphosphate, providing insight into the regulation of rRNA transcription. More broadly, the luciferase bioassay described here provides a general method for evaluating the ATP concentration available for biochemical processes in E. coli and potentially in other organisms. 10.1074/jbc.M311996200 IS - 9 JF - J. Biol. Chem. EP - 8268 TI - Relationship between Growth Rate and ATP Concentration in Escherichia coli: A BIOASSAY FOR AVAILABLE CELLULAR ATP VL - 279 SP - 8262 KW - atp KW - ecoli KW - growth KW - sampling AU - Schneider, David AU - Gourse, Richard PY - 2004/02/27/ UR - http://dx.doi.org/10.1074/jbc.M311996200 ER -