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J. Biol. Chem., Vol. 279, Issue 30, 31613-31621, July 23, 2004

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A Novel Fermentation/Respiration Switch Protein Regulated by Enzyme IIA^Glc in Escherichia coli^*

Byoung-Mo Koo, Mi-Jeong Yoon, Chang-Ro Lee, Tae-Wook Nam, Young-Jun Choe, Howard Jaffe, Alan Peterkofsky¶, and Yeong-Jae Seok||

From the Laboratory of Macromolecular Interactions, School of Biological Sciences and Institute of Microbiology, Seoul National University, Seoul 151-742, Korea, the NINDS, National Institutes of Health, Bethesda, Maryland 20892, and the ^¶Laboratory of Cell Biology, NHLBI, National Institutes of Health, Bethesda, Maryland 20892

The bacterial phosphoenolpyruvate:sugar phosphotransferase system regulates a variety of physiological processes as well as effecting sugar transport. The crr gene product (enzyme IIA^Glc (IIA^Glc)) mediates some of these regulatory phenomena. In this report, we characterize a novel IIA^Glc-binding protein from Escherichia coli extracts, discovered using ligand-fishing with surface plasmon resonance spectroscopy. This protein, which we named FrsA (fermentation/respiration switch protein), is the 47-kDa product of the yafA gene, previously denoted as "function unknown." FrsA forms a 1:1 complex specifically with the unphosphorylated form of IIA^Glc, with the highest affinity of any protein thus far shown to interact with IIA^Glc. Orthologs of FrsA have been found to exist only in facultative anaerobes belonging to the -proteobacterial group. Disruption of frsA increased cellular respiration on several sugars including glucose, while increased FrsA expression resulted in an increased fermentation rate on these sugars with the concomitant accumulation of mixed-acid fermentation products. These results suggest that IIA^Glc regulates the flux between respiration and fermentation pathways by sensing the available sugar species via a phosphorylation state-dependent interaction with FrsA.

Received for publication, May 6, 2004

^* This work was supported by a Korea Research Foundation Grant (KRF-2001-015-DS0057), in part by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Science & Technology (Grant MG02-0201-005-2-1-0), Republic of Korea, and by BK21 Research Fellowships from the Korean Ministry of Education and Human Resources Development (to B.-M. K., M.-J. Y., Y.-J. C., T.-W. N., and C.-R. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed. Tel.: 82-2-880-8827; Fax: 82-2-888-4911; E-mail: yjseok{at}plaza.snu.ac.kr.

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