alpha-Galactoside uptake in Rhizobium meliloti: isolation and characterization of agpA, a gene encoding a periplasmic binding protein required for melibiose and raffinose utilization.
Rhizobium meliloti can occupy at least two distinct ecological niches; it is found in the soil as a free-living saprophyte, and it also lives as a nitrogen-fixing intracellular symbiont in root nodules of alfalfa and related legumes. One approach to understanding how R. meliloti alters its physiology in order to become an integral part of a developing nodule is to identify and characterize genes that are differentially expressed by bacteria living inside nodules. We used a screen to identify genes under the control of the R. meliloti regulatory protein NodD3, SyrM, or SyrA. These regulatory proteins are expressed by bacteria growing inside the root nodule. One gene isolated in this screen was mapped to pSymB and displayed complex regulation. The gene was downregulated by the syrA gene product and also by glucose and succinate. This gene, referred to as agpA, encodes a periplasmic binding protein that is most similar to proteins from the periplasmic oligopeptide binding protein family. It is likely that AgpA binds alpha-galactosides, because alpha-galactosides induce the expression of agpA, and agpA mutants cannot utilize or transport these sugars. Activity of an agpA::TnphoA fusion was downregulated by SyrA. Because syrA is known to be expressed at high levels in intracellular symbiotic R. meliloti and at low levels in the free-living bacteria, we propose that AgpA may belong to the class of gene products whose expression decreases when R. meliloti becomes an intracellular symbiont.
Gage
D J
DJ
Department of Biological Sciences, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305-5020, USA. Gage@uconnvm.uconn.edu
Long
S R
SR
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001692:Biological Transport; D002236:Carbohydrate Conformation; D004268:DNA-Binding Proteins; D015536:Down-Regulation; D005697:Galactosides / Q000378:metabolism*; D005798:Genes, Bacterial; D008553:Melibiose / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D019897:Periplasm / Q000378:metabolism; D033902:Periplasmic Binding Proteins; D011887:Raffinose / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D016962:Sinorhizobium meliloti / Q000201:enzymology* / Q000235:genetics; D015534:Trans-Activators; D014157:Transcription Factors
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D005697:Galactosides; D026901:Membrane Transport Proteins; C061268:NodD3 protein, Sinorhizobium meliloti; D033902:Periplasmic Binding Proteins; C111069:SyrA protein, Rhizobium meliloti; C064814:SyrM protein, Rhizobium meliloti; D015534:Trans-Activators; D014157:Transcription Factors; C116035:agpA protein, Sinorhizobium meliloti; D008553:Melibiose; D011887:Raffinose