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Journal of Bacteriology, August 2003, p. 4539-4547, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4539-4547.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

DNA Array-Based Transcriptional Analysis of Asporogenous, Nonsolventogenic Clostridium acetobutylicum Strains SKO1 and M5

Christopher A. Tomas, Keith V. Alsaker, Hendrik P. J. Bonarius,{dagger} Wouter T. Hendriksen,{ddagger} He Yang, Jeffrey A. Beamish, Carlos J. Paredes, and Eleftherios T. Papoutsakis*

Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208

Received 16 December 2002/ Accepted 14 April 2003

The large-scale transcriptional program of two Clostridium acetobutylicum strains (SKO1 and M5) relative to that of the parent strain (wild type [WT]) was examined by using DNA microarrays. Glass DNA arrays containing a selected set of 1,019 genes (including all 178 pSOL1 genes) covering more than 25% of the whole genome were designed, constructed, and validated for data reliability. Strain SKO1, with an inactivated spo0A gene, displays an asporogenous, filamentous, and largely deficient solventogenic phenotype. SKO1 displays downregulation of all solvent formation genes, sigF, and carbohydrate metabolism genes (similar to genes expressed as part of the stationary-phase response in Bacillus subtilis) but also several electron transport genes. A major cluster of genes upregulated in SKO1 includes abrB, the genes from the major chemotaxis and motility operons, and glycosylation genes. Strain M5 displays an asporogenous and nonsolventogenic phenotype due to loss of the megaplasmid pSOL1, which contains all genes necessary for solvent formation. Therefore, M5 displays downregulation of all pSOL1 genes expressed in the WT. Notable among other genes expressed more highly in WT than in M5 were sigF, several two-component histidine kinases, spo0A, cheA, cheC, many stress response genes, fts family genes, DNA topoisomerase genes, and central-carbon metabolism genes. Genes expressed more highly in M5 include electron transport genes (but different from those downregulated in SKO1) and several motility and chemotaxis genes. Most of these expression patterns were consistent with phenotypic characteristics. Several of these expression patterns are new or different from what is known in B. subtilis and can be used to test a number of functional-genomic hypotheses.

* Corresponding author. Mailing address: Department of Chemical Engineering, Northwestern University, Evanston, IL 60208. Phone: (847) 491-7455. Fax: (847) 491-3728. E-mail: e-paps{at}northwestern.edu.

{dagger} Present address: DSM, 6401 JH Heerlen, The Netherlands.

{ddagger} Present address: Erasmus University, 3000 DR Rotterdam, The Netherlands.

Journal of Bacteriology, August 2003, p. 4539-4547, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4539-4547.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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