Wild nodules can be broken: proteomics of Frankia in field-collected root nodules
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With the genomes of three Frankia strains available, high-throughput proteomics methods can be used to reveal the set of proteins expressed by these bacteria in symbiosis with plants. A question we address is the degree to which the known genomes can be used to study proteomes of uncharacterized frankiae growing in field-collected root nodules. To this end, we have characterized the symbiotic proteomes of Frankia from three plant species, Alnus incana subsp. rugosa, Ceanothus americanus, and Elaeagnus angustifolia. Root nodule proteins were identified using two-dimensional liquid chromatography coupled to tandem mass spectrometry (LC MS/MS) of trypsin-digested protein samples. We identified 1300 Frankia proteins in A. incana nodules using the Frankia alni ACN14a genome and 1100 proteins from E. angustifolia nodules using the EAN1pec genome. In addition, over 100 proteins were identified from C. americanus nodules using a more limited one dimensional LC MS/MS analysis. Many of the most abundant proteins identified are involved in energy and nitrogen metabolism. The enzyme nitrogenase and the nitrogenase iron protein were among the most abundant proteins, reflecting the major process occurring in symbiosis. Several hundred plant proteins were also identified. We highlight the power of proteomics to uncover the physiology of symbiotic Frankia in the environment using heterologous genome information.
KeywordsFrankia Root nodules LC MS/MS Actinorhizal Proteomics Nitrogen fixation
MudPIT, LC MS/MS, and Mascot data analyses were performed by Kathy Stone, Mary LoPresti, and Tom Abbott, at the Mass Spectrometry and Protein Chemistry facility of the W.M. Keck Foundation Biotechnology Resource Laboratory at Yale University. We also thank Pascal LaPierre and Teddie Benson for assistance with bioinformatics.
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