Tyrosine nitration of flagellins: a response of Sinorhizobium meliloti to nitrosative stress.

Rhizobia are bacteria which can either live as free organisms in the soil or interact with plants of the legume family with, as a result, the formation of root organs called nodules in which differentiated endosymbiotic bacteria fix atmospheric nitrogen to the plant's benefit. In both lifestyles, rhizobia are exposed to nitric oxide (NO) which can be perceived as a signaling or toxic molecule. NO can act at the transcriptional level but can also modify proteins by S-nitrosylation of cysteine or nitration of tyrosine residues. However, only few molecular targets of NO have been described in bacteria and none of them have been characterized in rhizobia. Here we examined tyrosine nitration of Sinorhizobium meliloti proteins induced by NO. We found three tyrosine nitrated proteins in S. meliloti grown in free-living conditions, in response to an NO donor. Two nitroproteins were identified by mass spectrometry and correspond to flagellins A and B. We showed that one of the nitratable tyrosines is essential to flagellin function in motility.IMPORTANCE Rhizobia are found as free-living bacteria in the soil or in interaction with plants and are exposed to nitric oxide (NO) in both environments. NO is known to have many effects on animals, plants and bacteria where only few molecular targets of NO have been described yet. We identified flagellin A and B by mass spectrometry as tyrosine nitrated proteins in Sinorhizobium meliloti in vivo We also showed that one of the nitratable tyrosines is essential to flagellin function in motility. The results enhanced our understanding of NO effects on rhizobia. Identification of bacterial flagellin nitration opens a new possible role of NO in plant-microbe interactions.