title: Molecular characterization of the bet genes encoding glycine betaine synthesis in Sinorhizobium meliloti 102F34.
abstract: As a first step towards the elucidation of the molecular mechanisms responsible for the utilization of choline and glycine betaine (betaine) either as carbon and nitrogen sources or as osmoprotectants in Sinorhizobium meliloti, we selected a Tn5 mutant, LTS23-1020, which failed to grow on choline but grew on betaine. The mutant was deficient in choline dehydrogenase (CDH) activity, failed to oxidize [methyl-14C]choline to [methyl-14C]betaine, and did not use choline, but still used betaine, as an osmoprotectant. The Tn5 mutation in LTS23-1020 was complemented by plasmid pCHO34, isolated from a genomic bank of S. meliloti 102F34. Subcloning and DNA sequencing showed that pCHO34 harbours two ORFs which showed 60% and 57% identity with the Escherichia coli betB gene encoding betaine-aldehyde dehydrogenase (BADH) and betA gene encoding CDH, respectively. In addition to the homology with E. coli genes, the deduced sequence of the sinorhizobial BADH protein displays consensus sequences also found in plant BADHs. The deduced sequence of the sinorhizobial CDH protein shares only 21% identical residues with choline oxidase from Arthrobacter globiformis. The structural organization of the betBA genes in S. meliloti differs from that described in E. coli: (i) the two ORFs are separated by a 210 bp sequence containing inverted repeats resembling a putative rho-independent transcription terminator, and (ii) no sequence homologous to betT (high-affinity choline transport system) or betI (regulator) was found in the vicinity of the sinorhizobial betBA genes. Evidence is also presented that the S. meliloti betBA genes are not located on the megaplasmids.
authors.affiliation: Laboratoire de Biologie Végétale et Microbiologie, URA CNRS 1114, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
authors.forename: Jean-Alain
authors.initials: JA
authors.lastname: Pocard
authors.affiliation: Laboratoire de Biologie Végétale et Microbiologie, URA CNRS 1114, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Vincent
authors.affiliation: Laboratoire de Biologie Végétale et Microbiologie, URA CNRS 1114, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Boncompagni
authors.affiliation: Department of Agronomy and Range Science, University of California, Davis, CA 95616, USA.
authors.forename: Linda Tombras
authors.initials: LT
authors.lastname: Smith
authors.affiliation: Laboratoire de Biologie Végétale et Microbiologie, URA CNRS 1114, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Poggi
authors.affiliation: Laboratoire de Biologie Végétale et Microbiologie, URA CNRS 1114, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
authors.forename: Daniel Le
authors.initials: DL
authors.lastname: Rudulier
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics; D000445:Aldehyde Oxidoreductases / Q000235:genetics*; D000595:Amino Acid Sequence; D001622:Betaine / Q000031:analogs & derivatives / Q000378:metabolism*; D050728:Betaine-Aldehyde Dehydrogenase; D002794:Choline / Q000378:metabolism; D050545:Choline Dehydrogenase; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D000455:Medicago sativa / Q000382:microbiology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010084:Oxidation-Reduction; D018517:Plant Roots / Q000382:microbiology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016962:Sinorhizobium meliloti / Q000235:genetics*; D013559:Symbiosis; D014882:Water-Electrolyte Balance
chemical_list: D001622:Betaine; C026820:betaine aldehyde; D000429:Alcohol Oxidoreductases; D050545:Choline Dehydrogenase; D000445:Aldehyde Oxidoreductases; D050728:Betaine-Aldehyde Dehydrogenase; D002794:Choline