JB
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Other Versions of this Article:
JB.00340-08v1
190/14/5132    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Gardner, J. G.
Right arrow Articles by Escalante-Semerena, J. C.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gardner, J. G.
Right arrow Articles by Escalante-Semerena, J. C.

 Previous Article  |  Next Article 

Journal of Bacteriology, July 2008, p. 5132-5136, Vol. 190, No. 14
0021-9193/08/$08.00+0     doi:10.1128/JB.00340-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Biochemical and Mutational Analyses of AcuA, the Acetyltransferase Enzyme That Controls the Activity of the Acetyl Coenzyme A Synthetase (AcsA) in Bacillus subtilis{triangledown} ,§

Jeffrey G. Gardner and Jorge C. Escalante-Semerena*

Department of Bacteriology, University of Wisconsin, Madison, Wisconsin

Received 7 March 2008/ Accepted 7 May 2008

The acuABC genes of Bacillus subtilis comprise a putative posttranslational modification system. The AcuA protein is a member of the Gcn5-related N-acetyltransferase (GNAT) superfamily, the AcuC protein is a class I histone deacetylase, and the role of the AcuB protein is not known. AcuA controls the activity of acetyl coenzyme A synthetase (AcsA; EC 6.2.1.1) in this bacterium by acetylating residue Lys549. Here we report the kinetic analysis of wild-type and variant AcuA proteins. We contrived a genetic scheme for the identification of AcuA residues critical for activity. Changes at residues H177 and G187 completely inactivated AcuA and led to its rapid turnover. Changes at residues R42 and T169 were less severe. In vitro assay conditions were optimized, and an effective means of inactivating the enzyme was found. The basic kinetic parameters of wild-type and variant AcuA proteins were obtained and compared to those of eukaryotic GNATs. Insights into how the isolated mutations may exert their deleterious effect were investigated by using the crystal structure of an AcuA homolog.

* Corresponding author. Mailing address: Department of Bacteriology, University of Wisconsin, 6478 Microbial Sciences Building, 1550 Linden Dr., Madison, WI 53706-1567. Phone: (608) 262-7379. Fax: (608) 265-7909. E-mail: escalante{at}bact.wisc.edu

{triangledown} Published ahead of print on 16 May 2008.

§ Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, July 2008, p. 5132-5136, Vol. 190, No. 14
0021-9193/08/$08.00+0     doi:10.1128/JB.00340-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Appl. Environ. Microbiol. Infect. Immun. Eukaryot. Cell
Mol. Cell. Biol. J. Virol. Microbiol. Mol. Biol. Rev.
ALL ASM JOURNALS

Copyright © 2008 by the American Society for Microbiology. All rights reserved.