Role of feedback regulation of pantothenate kinase (CoaA) in control of coenzyme A levels in Escherichia coli.

Journal of Bacteriology
Charles O RockSuzanne Jackowski

Abstract

Pantothenate kinase (CoaA) is a key regulator of coenzyme A (CoA) biosynthesis in Escherichia coli, and its activity is controlled by feedback inhibition by CoA and its thioesters. The importance of feedback inhibition in the control of the intracellular CoA levels was tested by constructing three site-directed mutants of CoaA that were predicted to be feedback resistant based on the crystal structure of the CoaA-CoA binary complex. CoaA[R106A], CoaA[H177Q], and CoaA[F247V] were purified and shown to retain significant catalytic activity and be refractory to inhibition by CoA. CoaA[R106A] retained 50% of the catalytic activity of CoaA, whereas the CoaA[H177Q] and CoaA[F247V] mutants were less active. The importance of feedback control of CoaA to the intracellular CoA levels was assessed by expressing either CoaA or CoaA[R106A] in strain ANS3 [coaA15(Ts) panD2]. Cells expressing CoaA[R106A] had significantly higher levels of phosphorylated pantothenate-derived metabolites and CoA in vivo and excreted significantly more 4'-phosphopantetheine into the medium compared to cells expressing the wild-type protein. These data illustrate the key role of feedback regulation of pantothenate kinase in the control of intracellular CoA levels.

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Citations

Aug 12, 2009·The Journal of Biological Chemistry·Yuusuke YokoojiTadayuki Imanaka
Jul 16, 2009·Human Molecular Genetics·Zhihao WuBing Zhou
Mar 2, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Satyabrata DasM Vijayan
May 6, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·T Wubben, A D Mesecar
Jan 11, 2012·Bioscience, Biotechnology, and Biochemistry·Daisuke ItoShigeru Shigeoka
Jan 24, 2007·Proceedings of the National Academy of Sciences of the United States of America·Roberta LeonardiYong-Mei Zhang
Oct 27, 2009·Journal of Bacteriology·Masakazu TakagiShigeru Chohnan
Feb 6, 2007·Trends in Parasitology·Sylke Müller, Barbara Kappes
Jan 30, 2013·Biochemical Society Transactions·Haruyuki AtomiTadayuki Imanaka
Aug 12, 2014·Biochemical Society Transactions·Paola VencoValeria Tiranti
Jan 18, 2005·FEMS Microbiology Reviews·Patricia RenestoDidier Raoult
Jun 10, 2010·PLoS Computational Biology·Sidhartha GoyalNed S Wingreen
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Oct 3, 2003·Journal of Lipid Research·Gayathri RamaswamySuzanne Jackowski
Nov 19, 2004·The Journal of Biological Chemistry·Roberta LeonardiSuzanne Jackowski
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Jul 26, 2005·The Journal of Biological Chemistry·Yong-Mei ZhangSuzanne Jackowski
Oct 17, 2006·The Journal of Biological Chemistry·Rafal Jonczyk, Ulrich Genschel
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