The effector AvrRxo1 phosphorylates NAD in planta

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dc.contributor.authorShidore, Tejaen
dc.contributor.authorBroeckling, Corey D.en
dc.contributor.authorKirkwood, Jay S.en
dc.contributor.authorLong, John J.en
dc.contributor.authorMiao, Jiaminen
dc.contributor.authorZhao, Bingyu Y.en
dc.contributor.authorLeach, Jan E.en
dc.contributor.authorTriplett, Lindsay R.en
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2018-12-04T18:42:14Zen
dc.date.available2018-12-04T18:42:14Zen
dc.date.issued2017-06-19en
dc.description.abstractGram-negative bacterial pathogens of plants and animals employ type III secreted effectors to suppress innate immunity. Most characterized effectors work through modification of host proteins or transcriptional regulators, although a few are known to modify small molecule targets. The Xanthomonas type III secreted avirulence factor AvrRxo1 is a structural homolog of the zeta toxin family of sugar-nucleotide kinases that suppresses bacterial growth. AvrRxo1 was recently reported to phosphorylate the central metabolite and signaling molecule NAD in vitro, suggesting that the effector might enhance bacterial virulence on plants through manipulation of primary metabolic pathways. In this study, we determine that AvrRxo1 phosphorylates NAD in planta, and that its kinase catalytic sites are necessary for its toxic and resistance-triggering phenotypes. A global metabolomics approach was used to independently identify 3'-NADP as the sole detectable product of AvrRxo1 expression in yeast and bacteria, and NAD kinase activity was confirmed in vitro. 3'-NADP accumulated upon transient expression of AvrRxo1 in Nicotiana benthamiana and in rice leaves infected with avrRxo1-expressing strains of X. oryzae. Mutation of the catalytic aspartic acid residue D193 abolished AvrRxo1 kinase activity and several phenotypes of AvrRxo1, including toxicity in yeast, bacteria, and plants, suppression of the flg22-triggered ROS burst, and ability to trigger an R gene-mediated hypersensitive response. A mutation in the Walker A ATPbinding motif abolished the toxicity of AvrRxo1, but did not abolish the 3'-NADP production, virulence enhancement, ROS suppression, or HR-triggering phenotypes of AvrRxo1. These results demonstrate that a type III effector targets the central metabolite and redox carrier NAD in planta, and that this catalytic activity is required for toxicity and suppression of the ROS burst.en
dc.description.sponsorshipThis project was funded by the Agriculture and Food Research Initiative (nifa.usda.gov) competitive grant no. 2014-67013-21564 of the USDA National Institute of Food and Agriculture to LRT and JEL. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.en
dc.format.extent22 pagesen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1371/journal.ppat.1006442en
dc.identifier.eissn1553-7374en
dc.identifier.issn1553-7366en
dc.identifier.issue6en
dc.identifier.othere1006442en
dc.identifier.pmid28628666en
dc.identifier.urihttp://hdl.handle.net/10919/86215en
dc.identifier.volume13en
dc.language.isoenen
dc.publisherPLOSen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectnicotinamide-nucleotide transhydrogenaseen
dc.subjectt4 polynucleotide kinaseen
dc.subjectmalic enzymeen
dc.subjectcrystal-structureen
dc.subjectoryzaeen
dc.subjectriceen
dc.subjectresistanceen
dc.subjectinhibitorsen
dc.subjectimmunityen
dc.subjectmaizeen
dc.titleThe effector AvrRxo1 phosphorylates NAD in plantaen
dc.title.serialPLOS Pathogensen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
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