In vitro sulfotransferase activity of NoeE, a nodulation protein of Rhizobium sp. NGR234.
Soil bacteria of the genera Azorhizobium, Bradyrhizobium, and Rhizobium liberate morphogenetic lipochitin-oligosaccharides (Nod factors) into legume rhizospheres. Nod factors, which are synthesized by the products of rhizobial nodulation (nod) genes, vary in core length as well as in the number and type of substitutions. In Rhizobium sp. NGR234, the N-acylated pentamers of N-acetyl-D-glucosamine carry an O-methylfucose group on the reducing terminus that is substituted, on a mutually exclusive basis, with either an acetyl or a sulfuryl group. A sulfotransferase encoded by noeE is required for adjunction of activated sulfate donated by 3'-phosphoadenosine 5'-phosphosulfate (PAPS). Here we show that when expressed in NGR234 cured of its symbiotic plasmid (= ANU265) or when purified as a fusion protein (MBP-NoeE), NoeE transfers sulfate from PAPS to fucosylated lipochitin-oligosaccharides. Enzyme assays showed that sulfotransferase activity is dependent on the presence of an acyl group (stearic and vaccenic acids were tested) since no activity was detected when fucosylated oligochitins (oligomers of two to six N-acetyl-D-glucosamine units) were used as substrates. Thus, NoeE is unique in that it is the only characterized sulfotransferase that is specific for fucosylated Nod factors. It probably acts after NodA, which acylates the amino-sugar backbone.
Quesada-Vincens
D
D
Laboratoire de Biologie Moléculaire des Plantes Supérieures, Université de Genève, Switzerland.
Hanin
M
M
Broughton
W J
WJ
Jabbouri
S
S
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000378:metabolism*; D001483:Base Sequence; D002240:Carbohydrate Sequence; D002686:Chitin / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004926:Escherichia coli; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012231:Rhizobium / Q000201:enzymology*; D015238:Sulfotransferases / Q000378:metabolism*
D001426:Bacterial Proteins; D009838:Oligodeoxyribonucleotides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D002686:Chitin; C107979:NoeE protein, Rhizobium sp.; D015238:Sulfotransferases