In vitro sulfotransferase activity of NodH, a nodulation protein of Rhizobium meliloti required for host-specific nodulation.
Early stages of nodulation involve the exchange of signals between the bacterium and the host plant. Bacterial nodulation (nod) genes are required for Rhizobium spp. to synthesize lipooligosaccharide morphogens, termed Nod factors. The common nod genes encode enzymes that synthesize the factor core structure, which is modified by host-specific gene products. Here we show direct in vitro evidence that Rhizobium meliloti NodH, a host-specific nodulation gene, catalyzes the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to the terminal 6-O position of Nod factors, and we show substrate requirements for the reaction. Our results indicate that polymerization of the chitooligosaccharide backbone likely precedes sulfation and that sulfation is not absolutely dependent on the presence or the particular structure of the N-acyl modification. NodH sulfation provides a tool for the enzymatic in vitro synthesis of novel Nod factors, or putative Nod factors intermediates, with high specific activity.
Ehrhardt
D W
DW
Gilbert Laboratory, Howard Hughes Medical Institute, Stanford University, California 94305-5020, USA.
Atkinson
E M
EM
Faull
K F
KF
Freedberg
D I
DI
Sutherlin
D P
DP
Armstrong
R
R
Long
S R
SR
D001426:Bacterial Proteins; D002240:Carbohydrate Sequence; D002686:Chitin / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation; D009844:Oligosaccharides / Q000378:metabolism; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism; D018517:Plant Roots / Q000382:microbiology*; D011994:Recombinant Proteins / Q000378:metabolism; D016962:Sinorhizobium meliloti / Q000201:enzymology* / Q000235:genetics; D013045:Species Specificity; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism*; D013463:Sulfuric Acid Esters / Q000378:metabolism; D013559:Symbiosis
D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D013463:Sulfuric Acid Esters; D002686:Chitin; D010724:Phosphoadenosine Phosphosulfate; C092976:NodH protein, Sinorhizobium meliloti; D015238:Sulfotransferases