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Journal of Bacteriology, September 2005, p. 5901-5909, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.5901-5909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

ABC Transporter for Corrinoids in Halobacterium sp. Strain NRC-1{dagger}

Jesse D. Woodson, April A. Reynolds, and Jorge C. Escalante-Semerena*

Department of Bacteriology, University of Wisconsin—Madison, Madison, Wisconsin

Received 1 March 2005/ Accepted 26 May 2005

We report evidence for the existence of a putative ABC transporter for corrinoid utilization in the extremely halophilic archaeon Halobacterium sp. strain NRC-1. Results from genetic and nutritional analyses of Halobacterium showed that mutants with lesions in open reading frames (ORFs) Vng1370G, Vng1371Gm, and Vng1369G required a 105-fold higher concentration of cobalamin for growth than the wild-type or parent strain. The data support the conclusion that these ORFs encode orthologs of the bacterial cobalamin ABC transporter permease (btuC; Vng1370G), ATPase (btuD; Vng1371Gm), and substrate-binding protein (btuF; Vng1369G) components. Mutations in the Vng1370G, Vng1371Gm, and Vng1369G genes were epistatic, consistent with the hypothesis that their products work together to accomplish the same function. Extracts of btuF mutant strains grown in the presence of cobalamin did not contain any cobalamin molecules detectable by a sensitive bioassay, whereas btuCD mutant strain extracts did. The data are consistent with the hypothesis that the BtuF protein is exported to the extracellular side of the cell membrane, where it can bind cobalamin in the absence of BtuC and BtuD. Our data also provide evidence for the regulation of corrinoid transport and biosynthesis. Halobacterium synthesized cobalamin in a chemically defined medium lacking corrinoid precursors. To the best of our knowledge, this is the first genetic analysis of an archaeal corrinoid transport system.


* Corresponding author. Mailing address: Department of Bacteriology, University of Wisconsin, 144A Enzyme Institute, 1710 University Avenue, Madison, WI 53726-4087. Phone: (608) 262-7379. Fax: (608) 265-7909. E-mail: escalante{at}bact.wisc.edu.

{dagger} We dedicate this work to the memory of Robert Kadner, a pioneer in the field of corrinoid transport, a good friend and colleague.


Journal of Bacteriology, September 2005, p. 5901-5909, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.5901-5909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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