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Journal of Bacteriology, October 1999, p. 6238-6246, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Genetic and Biochemical Characterization of a High-Affinity
Betaine Uptake System (BusA) in Lactococcus lactis
Reveals a New Functional Organization within Bacterial
ABC Transporters
David
Obis,1
Alain
Guillot,1
Jean-Claude
Gripon,1
Pierre
Renault,2
Alexander
Bolotin,2 and
Michel-Yves
Mistou1,*
Unité de Biochimie et Structure des
Protéines1 and Unité de
Génétique Microbienne,2 Institut
National de la Recherche Agronomique, 78352 Jouy-en-Josas Cedex, France
Received 20 April 1999/Accepted 30 July 1999
The cytoplasmic accumulation of exogenous betaine stimulates the
growth of Lactococcus lactis cultivated under hyperosmotic conditions. We report that L. lactis possesses a single
betaine transport system that belongs to the ATP-binding cassette
(ABC) superfamily of transporters. Through transposon mutagenesis,
a mutant deficient in betaine transport was isolated. We
identified two genes, busAA and busAB, grouped
in an operon, busA (betaine uptake system).
The transcription of busA is strongly regulated by the
external osmolality of the medium. The busAA gene codes for
the ATP-binding protein. busAB encodes a 573-residue
polypeptide which presents two striking features: (i) a fusion between
the regions encoding the transmembrane domain (TMD) and the
substrate-binding domain (SBD) and (ii) a swapping of the SBD
subdomains when compared to the Bacillus subtilis
betaine-binding protein, OpuAC. BusA of L. lactis displays
a high affinity towards betaine (Km = 1.7 µM)
and is an osmosensor whose activity is tightly regulated by external
osmolality, leading the betaine uptake capacity of
L. lactis to be under dual control at the biochemical and
genetic levels. A protein presenting the characteristics
predicted for BusAB was detected in the membrane fraction of L. lactis. The fusion between the TMD and the SBD is the first
example of a new organization within prokaryotic ABC transporters.
*
Corresponding author. Mailing address: Unité de
Biochimie et Structure des Protéines, Institut National de la
Recherche Agronomique, 78352 Jouy-en-Josas Cedex, France.
Phone: 33-1-34-65-22-62. Fax: 33-1-34-65-21-63. E-mail:
mistou{at}jouy.inra.fr.
Journal of Bacteriology, October 1999, p. 6238-6246, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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