JBC Transcription and Nuclear Factor Monoclonals

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J Biol Chem, Vol. 275, Issue 2, 1377-1383, January 14, 2000

Pantothenate Kinase Regulation of the Intracellular Concentration of Coenzyme A*

Charles O. RockDagger §, Robert B. CalderDagger , Mohammad A. KarimDagger , and Suzanne JackowskiDagger §

From the Dagger  Department of Biochemistry, St. Jude Children's Research Hospital, Memphis, Tennessee 38105 and the § Department of Biochemistry, University of Tennessee, Memphis, Tennessee 38163

Pantothenate kinase (PanK) is the key regulatory enzyme in the CoA biosynthetic pathway in bacteria and is thought to play a similar role in mammalian cells. We examined this hypothesis by identifying and characterizing two murine cDNAs that encoded PanK. The two cDNAs were predicted to arise from alternate splicing of the same gene to yield different mRNAs that encode two isoforms (mPanK1alpha and mPanK1beta ) with distinct amino termini. The predicted protein sequence of mPanK1 was not related to bacterial PanK but exhibited significant similarity to Aspergillus nidulans PanK. mPanK1alpha was most highly expressed in heart and kidney, whereas mPanK1beta mRNA was detected primarily in liver and kidney. Pantothenate was the most abundant pathway component (42.8%) in normal cells providing clear evidence that pantothenate phosphorylation was a rate-controlling step in CoA biosynthesis. Enhanced mPanK1beta expression eliminated the intracellular pantothenate pool and triggered a 13-fold increase in intracellular CoA content. mPanK1beta activity in vitro was stimulated by CoA and strongly inhibited by acetyl-CoA illustrating that differential modulation of mPanK1beta activity by pathway end products also contributed to the management of CoA levels. These data support the concept that the expression and/or activity of PanK is a determining factor in the physiological regulation of the intracellular CoA concentration.

* This work was supported by National Institutes of Health Grants GM 45737 (to S. J.) and GM34496 (to C. O. R.), Cancer Center (CORE) Support Grant CA 21765, and the American Lebanese Syrian Associated Charities.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF200357.

To whom correspondence should be addressed: Biochemistry Dept., 332 N. Lauderdale, Memphis, TN 38105-2794. Tel.: 901-495-3494; Fax: 901-525-8025; E-mail: suzanne.jackowski@stjude.org.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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