PMID: 9537383Apr 16, 1998Paper

Novel assay reveals multiple pathways regulating stress-induced accumulations of inorganic polyphosphate in Escherichia coli

Journal of Bacteriology
D Ault-RichéA Kornberg

Abstract

A major impediment to understanding the biological roles of inorganic polyphosphate (polyP) has been the lack of sensitive definitive methods to extract and quantitate cellular polyP. We show that polyP recovered in extracts from cells lysed with guanidinium isothiocynate can be bound to silicate glass and quantitatively measured by a two-enzyme assay: polyP is first converted to ATP by polyP kinase, and the ATP is hydrolyzed by luciferase to generate light. This nonradioactive method can detect picomolar amounts of phosphate residues in polyP per milligram of extracted protein. A simplified procedure for preparing polyP synthesized by polyP kinase is also described. Using the new assay, we found that bacteria subjected to nutritional or osmotic stress in a rich medium or to nitrogen exhaustion had large and dynamic accumulations of polyP. By contrast, carbon exhaustion, changes in pH, temperature upshifts, and oxidative stress had no effect on polyP levels. Analysis of Escherichia coli mutants revealed that polyP accumulation depends on several regulatory genes, glnD (NtrC), rpoS, relA, and phoB.

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Citations

Jun 29, 2000·Annual Review of Biochemistry·A KornbergD Ault-Riché
Aug 10, 2000·Proceedings of the National Academy of Sciences of the United States of America·M H RashidA Kornberg
Aug 31, 2000·Applied and Environmental Microbiology·J W McGrath, J P Quinn
Nov 4, 2000·Molecular Plant-microbe Interactions : MPMI·P M RiccilloO M Aguilar
Oct 24, 2002·The Journal of Biological Chemistry·Claudia O RodriguesRoberto Docampo
Dec 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Haiyu ZhangArthur Kornberg
Feb 11, 2004·The Journal of Biological Chemistry·Bernard LakayeLucien Bettendorff
Apr 1, 2006·Applied Microbiology and Biotechnology·Alexandra D HollandMary E Lidstrom
Aug 23, 2006·Journal of Bacteriology·Sofiane GhorbelMarie-Joelle Virolle
Aug 7, 2008·Journal of Food Protection·Jeremy A ObritschLloyd B Bullerman
Jun 23, 2009·FEMS Microbiology Letters·Kristina Jonas, Ojar Melefors
Aug 16, 2011·The Journal of Biological Chemistry·David DelvauxLucien Bettendorff
Sep 13, 2011·Antonie van Leeuwenhoek·Jörg BrockHeide N Schulz-Vogt
Apr 6, 2012·International Journal of Hematology·James H Morrissey
Nov 6, 2012·Journal of Bacteriology·Chuan WangQian Gao
Apr 23, 2014·Proceedings of the National Academy of Sciences of the United States of America·Patrick MartinBenjamin A S Van Mooy
Apr 25, 2015·Arteriosclerosis, Thrombosis, and Vascular Biology·Stephanie A Smith, James H Morrissey
Nov 18, 2015·Enzyme Research·Paola R BeassoniAngela T Lisa
Jan 24, 2017·Nature Microbiology·Jan-Ulrik DahlUrsula Jakob
Sep 27, 2002·Applied and Environmental Microbiology·Katherine D McMahonJay D Keasling
Jul 31, 2002·Applied and Environmental Microbiology·Tomohiro MorohoshiAkio Kuroda
Jul 16, 2002·Infection and Immunity·Wen ChenHoward K Kuramitsu
Sep 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·Lihong WangRichard A Roth
Nov 5, 2004·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Felix A RuizRoberto Docampo
Feb 11, 2005·Proceedings of the National Academy of Sciences of the United States of America·Haiyu ZhangArthur Kornberg
Jan 23, 2008·Journal of Fluorescence·Roozbeh Aschar-SobbiEvgeny Pavlov

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