Tyrosine sulfation in a Gram-negative bacterium

Tyrosine sulfation, a well-characterized post-translation modification in eukaryotes, has not previously been reported in prokaryotes. Here, we demonstrate that the RaxST protein from the Gram-negative bacterium, Xanthomonas oryzae pv. oryzae, is a tyrosine sulfotransferase. We used a newly developed sulfotransferase assay and ultraviolet photodissociation mass spectrometry to demonstrate that RaxST catalyses sulfation of tyrosine 22 of the Xoo Ax21 (activator of XA21-mediated immunity) protein. These results demonstrate a previously undescribed post-translational modification in a prokaryotic species with implications for studies of host immune responses and bacterial cell-cell communication systems.

[1]  H. Glatt,et al.  Differential activation of promutagens by alloenzymes of human sulfotransferase 1A2 expressed in Salmonella typhimurium. , 2002, Pharmacogenetics.

[2]  D. Alessi,et al.  The nuts and bolts of AGC protein kinases , 2010, Nature Reviews Molecular Cell Biology.

[3]  R. Glockshuber,et al.  DsbL and DsbI form a specific dithiol oxidase system for periplasmic arylsulfate sulfotransferase in uropathogenic Escherichia coli. , 2008, Journal of molecular biology.

[4]  D. H. Keating,et al.  Sinorhizobium meliloti SyrA Mediates the Transcriptional Regulation of Genes Involved in Lipopolysaccharide Sulfation and Exopolysaccharide Biosynthesis , 2007, Journal of bacteriology.

[5]  J. M. Dow,et al.  The Ax21 Protein Is a Cell-Cell Signal That Regulates Virulence in the Nosocomial Pathogen Stenotrophomonas maltophilia , 2011, Journal of bacteriology.

[6]  Mei Li,et al.  Crystal structure of human sulfotransferase SULT1A3 in complex with dopamine and 3'-phosphoadenosine 5'-phosphate. , 2005, Biochemical and biophysical research communications.

[7]  Joseph Sodroski,et al.  Tyrosine Sulfation of the Amino Terminus of CCR5 Facilitates HIV-1 Entry , 1999, Cell.

[8]  P. Ronald,et al.  Small Protein-Mediated Quorum Sensing in a Gram-Negative Bacterium , 2011, PloS one.

[9]  A. Venter,et al.  Journal of The American Society for Mass Spectrometry , 2005, Journal of the American Society for Mass Spectrometry.

[10]  J. Brodbelt.  Shedding Light on the Frontier of Photodissociation , 2011, Journal of the American Society for Mass Spectrometry.

[11]  Y. Matsubayashi,et al.  An LRR Receptor Kinase Involved in Perception of a Peptide Plant Hormone, Phytosulfokine , 2002, Science.

[12]  P. Ronald,et al.  A Type I–Secreted, Sulfated Peptide Triggers XA21-Mediated Innate Immunity , 2009, Science.

[13]  P. Ronald,et al.  Secretion, modification, and regulation of Ax21. , 2011, Current opinion in microbiology.

[14]  P. Ronald,et al.  Bacterial genes involved in type I secretion and sulfation are required to elicit the rice Xa21-mediated innate immune response. , 2004, Molecular plant-microbe interactions : MPMI.

[15]  Glen P. Jackson,et al.  Metastable Atom-Activated Dissociation Mass Spectrometry of Phosphorylated and Sulfonated Peptides in Negative Ion Mode , 2011, Journal of the American Society for Mass Spectrometry.

[16]  Spencer J. Williams,et al.  Discovery of sulfated metabolites in mycobacteria with a genetic and mass spectrometric approach , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Ronald,et al.  Unique characteristics of Xanthomonas oryzae pv. oryzae AvrXa21 and implications for plant innate immunity , 2006, Proceedings of the National Academy of Sciences.

[18]  P. Ronald,et al.  The Xanthomonas oryzae pv.◊oryzae raxP and raxQ genes encode an ATP sulphurylase and adenosine‐5′‐phosphosulphate kinase that are required for AvrXa21 avirulence activity , 2002, Molecular microbiology.

[19]  Ying Sun,et al.  A conserved HIV gp120 glycoprotein structure involved in chemokine receptor binding. , 1998, Science.

[20]  I. H. Segel,et al.  Adenosine 5'-phosphosulfate kinase from Penicillium chrysogenum. Purification and kinetic characterization. , 1984, The Journal of biological chemistry.

[21]  D. Sutherlin,et al.  In vitro sulfotransferase activity of NodH, a nodulation protein of Rhizobium meliloti required for host-specific nodulation , 1995, Journal of bacteriology.

[22]  M. Cadene,et al.  Tyrosine sulfation of CCR5 N-terminal peptide by tyrosylprotein sulfotransferases 1 and 2 follows a discrete pattern and temporal sequence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. Brodbelt,et al.  193‐nm photodissociation of singly and multiply charged peptide anions for acidic proteome characterization , 2011, Proteomics.

[24]  K. Moore.  Protein tyrosine sulfation: A critical posttranslation modification in plants and animals , 2009, Proceedings of the National Academy of Sciences.

[25]  M. Westphall,et al.  Implementing Photodissociation in an Orbitrap Mass Spectrometer , 2011, Journal of the American Society for Mass Spectrometry.

[26]  D. Boutz,et al.  Ultrafast ultraviolet photodissociation at 193 nm and its applicability to proteomic workflows. , 2010, Journal of proteome research.

[27]  M. Shoham,et al.  Tyrosine sulfation: an increasingly recognised post-translational modification of secreted proteins. , 2009, New biotechnology.

[28]  P. Lerouge,et al.  Molecular basis of symbiotic host specificity in rhizobium meliloti: nodH and nodPQ genes encode the sulfation of lipo-oligosaccharide signals , 1991, Cell.