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NodS is a 5-adenosyl-L-methionine-dependent methyltransferase that methylates chitooligosaccharides deacetylated at the nonreducing end

Danny Geelen (UGent) , Barbara Leyman (UGent) , Peter Mergaert (UGent) , Klaus Klarskov, Marc Van Montagu (UGent) , Roberto Geremia (UGent) and Marcella Holsters (UGent)
(1995) MOLECULAR MICROBIOLOGY. 17(2). p.387-397
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Abstract
In response to phenolic compounds exuded by the host plant, symbiotic Rhizobium bacteria produce signal molecules (Nod factors), consisting of lipochitooligosaccharides with strain-specific substitutions. In Azorhizobium caulinodans strain ORS571 these modifications are an O-arabinosyl group, an O-carbamoyl group, and an IV-methyl group. Several lines of evidence indicate that the nodS gene located in the nodABCSUIJ operon is implicated in the methylation of Nod factors, Previously we have shown that NodS is an S-adenosyl-L-methionine (SAM)-binding protein, essential for the L-[H-3-methyl]-methionine labelling of ORS571 Nod factors in vivo. Here, we present an in vitro assay showing that NodS from either A. caulinodans or Rhizobium species NGR234 methylates end-deacetylated chitooligosaccharides, using [H-3-methyl]-SAM as a methyl donor. The enzymatic and SAM-binding activity were correlated with the nodS gene and localized within the soluble protein fraction. The A. caulinodans nodS gene was expressed in Escherichia coil and a glutathione-S-transferase-NodS fusion protein purified. This protein bound SAM and could methylate end-deacetylated chitooligosaccharides, but could not fully methylate acetylated chitooligosaccharides or unmethylated lipo-chitooligosaccharides. These data implicate that the methylation step in the biosynthesis pathway of ORS571 Nod factors occurs after deacetylation and prior to acylation of the chitooligosaccharides.
Keywords
SESBANIA-ROSTRATA, NODULATION SIGNALS, STRAIN NGR234, BRADYRHIZOBIUM-JAPONICUM, AZORHIZOBIUM-CAULINODANS, LIPO-OLIGOSACCHARIDE SIGNALS, RHIZOBIUM-MELILOTI, HOST SPECIFICITY, GENES, IDENTIFICATION

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Chicago
Geelen, Danny, Barbara Leyman, Peter Mergaert, Klaus Klarskov, Marc Van Montagu, Roberto Geremia, and Marcella Holsters. 1995. “NodS Is a 5-adenosyl-L-methionine-dependent Methyltransferase That Methylates Chitooligosaccharides Deacetylated at the Nonreducing End.” Molecular Microbiology 17 (2): 387–397.
APA
Geelen, D., Leyman, B., Mergaert, P., Klarskov, K., Van Montagu, M., Geremia, R., & Holsters, M. (1995). NodS is a 5-adenosyl-L-methionine-dependent methyltransferase that methylates chitooligosaccharides deacetylated at the nonreducing end. MOLECULAR MICROBIOLOGY, 17(2), 387–397.
Vancouver
1.
Geelen D, Leyman B, Mergaert P, Klarskov K, Van Montagu M, Geremia R, et al. NodS is a 5-adenosyl-L-methionine-dependent methyltransferase that methylates chitooligosaccharides deacetylated at the nonreducing end. MOLECULAR MICROBIOLOGY. 1995;17(2):387–97.
MLA
Geelen, Danny, Barbara Leyman, Peter Mergaert, et al. “NodS Is a 5-adenosyl-L-methionine-dependent Methyltransferase That Methylates Chitooligosaccharides Deacetylated at the Nonreducing End.” MOLECULAR MICROBIOLOGY 17.2 (1995): 387–397. Print.
@article{195165,
  abstract     = {In response to phenolic compounds exuded by the host plant, symbiotic Rhizobium bacteria produce signal molecules (Nod factors), consisting of lipochitooligosaccharides with strain-specific substitutions. In Azorhizobium caulinodans strain ORS571 these modifications are an O-arabinosyl group, an O-carbamoyl group, and an IV-methyl group. Several lines of evidence indicate that the nodS gene located in the nodABCSUIJ operon is implicated in the methylation of Nod factors, Previously we have shown that NodS is an S-adenosyl-L-methionine (SAM)-binding protein, essential for the L-[H-3-methyl]-methionine labelling of ORS571 Nod factors in vivo. Here, we present an in vitro assay showing that NodS from either A. caulinodans or Rhizobium species NGR234 methylates end-deacetylated chitooligosaccharides, using [H-3-methyl]-SAM as a methyl donor. The enzymatic and SAM-binding activity were correlated with the nodS gene and localized within the soluble protein fraction. The A. caulinodans nodS gene was expressed in Escherichia coil and a glutathione-S-transferase-NodS fusion protein purified. This protein bound SAM and could methylate end-deacetylated chitooligosaccharides, but could not fully methylate acetylated chitooligosaccharides or unmethylated lipo-chitooligosaccharides. These data implicate that the methylation step in the biosynthesis pathway of ORS571 Nod factors occurs after deacetylation and prior to acylation of the chitooligosaccharides.},
  author       = {Geelen, Danny and Leyman, Barbara and Mergaert, Peter and Klarskov, Klaus and Van Montagu, Marc and Geremia, Roberto and Holsters, Marcella},
  issn         = {0950-382X},
  journal      = {MOLECULAR MICROBIOLOGY},
  keyword      = {SESBANIA-ROSTRATA,NODULATION SIGNALS,STRAIN NGR234,BRADYRHIZOBIUM-JAPONICUM,AZORHIZOBIUM-CAULINODANS,LIPO-OLIGOSACCHARIDE SIGNALS,RHIZOBIUM-MELILOTI,HOST SPECIFICITY,GENES,IDENTIFICATION},
  language     = {eng},
  number       = {2},
  pages        = {387--397},
  title        = {NodS is a 5-adenosyl-L-methionine-dependent methyltransferase that methylates chitooligosaccharides deacetylated at the nonreducing end},
  url          = {http://dx.doi.org/10.1111/j.1365-2958.1995.mmi\_17020387.x},
  volume       = {17},
  year         = {1995},
}

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