A glutamate/glutamine/aspartate/asparagine transport operon in Rhodobacter capsulatus.
A mutant of Rhodobacter capsulatus was identified in which an operon encoding a binding-protein-dependent transporter was interrupted by Tn5 transposition. Cloning and sequence analysis of the wild-type operon revealed a four-gene cluster with similarities to genes encoding periplasmic binding proteins (BztA), integral membrane proteins (BztB and BztC), and ATP-binding proteins (BztD). To assess the function of this putative binding-protein-dependent transport system, a mutant was constructed in which most of the bztABCD operon was deleted and replaced by an antibiotic-resistance marker. The deletion mutant grew more slowly than the wild type in NH4(+)-free medium supplemented by glutamate, glutamine, aspartate or asparagine; it was resistant to toxic analogues of Glu, Asp, and Asn at concentrations that inhibited growth of the wild type; and it was defective in the uptake of Glu, Gln, and Asp. A complementing plasmid containing the wildtype copy of bztABCD was able to rescue all the mutant phenotypes. Taken together, these results indicate that the proteins encoded by bztABCD are active in the uptake of Glu, Gln, Asp, and Asn. In addition, competition experiments, in which the ability of each of the four amino acids to compete for the transport of one another was examined, demonstrated that all four substrates share at least one component of this transport system.
Zheng
S
S
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637, USA.
Haselkorn
R
R
D000595:Amino Acid Sequence; D026905:Amino Acid Transport Systems; D001216:Asparagine / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018698:Glutamic Acid / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D008969:Molecular Sequence Data; D009876:Operon; D016354:Rhodobacter capsulatus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D026905:Amino Acid Transport Systems; D002352:Carrier Proteins; D005973:Glutamine; D001224:Aspartic Acid; D018698:Glutamic Acid; D001216:Asparagine