Genetic and Proteomic Analyses of a Xanthomonas campestris pv. campestris purC Mutant Deficient in Purine Biosynthesis and Virulence

Zhihui Yuan; Li Wang; Shutao Sun; Yao Wu; Wei Qian

doi:10.1016/j.jgg.2013.05.003

Volume 40 Issue 9

Sep. 2013

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Genetic and Proteomic Analyses of a Xanthomonas campestris pv. campestris purC Mutant Deficient in Purine Biosynthesis and Virulence

doi: 10.1016/j.jgg.2013.05.003

Zhihui Yuan^{a, b, #},
Li Wang^{a, #},
Shutao Sun^c,
Yao Wu^a,
Wei Qian^a

a
State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
b
University of Chinese Academy of Sciences, Beijing 100049, China
c
Center of Core Facility, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China

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Corresponding author: E-mail address: qianw@im.ac.cn (Wei Qian)
Received Date: 2013-03-11
Accepted Date: 2013-05-23
Rev Recd Date: 2013-05-21

Available Online: 2013-06-01

Publish Date: 2013-09-20

Abstract

Abstract

Bacterial proliferation in hosts requires activation of a number of housekeeping pathways, including purine de novo biosynthesis. Although inactivation of purine biosynthesis genes can attenuate virulence, it is unclear which biochemical or virulence factors are associated with the purine biosynthesis pathway in vivo. We report that inactivation of purC, a gene encoding phosphoribosylaminoimidazole-succinocarboxamide synthase, caused complete loss of virulence inXanthomonas campestris pv. campestris, the causal agent of black rot disease of cruciferous plants. The purC mutant was a purine auxotroph; it could not grow on minimal medium, whereas addition of purine derivatives, such as hypoxanthine or adenine plus guanine, restored growth of the mutant. The purC mutation also significantly enhanced the production of an unknown purine synthesis associated pigment and extracellular polysaccharides by the bacterium. In addition, comparative proteomic analyses of bacteria grown on rich and minimal media revealed that the purC mutation affected the expression levels of diverse proteins involved in purine and pyrimidine synthesis, carbon and energy metabolisms, iron uptake, proteolysis, protein secretion, and signal transduction. These results provided clues to understanding the contributions of purine synthesis to bacterial virulence and interactions with host immune systems.
- Purine biosynthesis,
- Virulence,
- Proteomics

These authors contributed equally to this work.

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References

[1]	Alcantara, R.B., Read, R.D., Valderas, M.W. et al. Infect. Immun., 72 (2004),pp. 4911-4917
[2]	Andrews, S.C., Robinson, A.K., Rodriguez-Quinones, F. Bacterial iron homeostasis FEMS Microbiol. Rev., 27 (2003),pp. 215-237
[3]	Beyer, N.H., Roepstorff, P., Hammer, K. et al. Proteomics, 3 (2003),pp. 786-797
[4]	Bigot, A., Botton, E., Dubail, I. et al. Appl. Environ. Microbiol., 72 (2006),pp. 6623-6631
[5]	Bingel-Erlenmeyer, R., Kohler, R., Kramer, G. et al. A peptide deformylase-ribosome complex reveals mechanism of nascent chain processing Nature, 452 (2008),pp. 108-111
[6]	Buttner, D., Bonas, U. FEMS Microbiol. Rev., 34 (2010),pp. 107-133
[7]	Cattoir, V., Narasimhan, G., Skurnik, D. et al. MBio, 3 (2013),pp. e00410-e00412
[8]	Chatterjee, S., Sonti, R.V. Can. J. Microbiol., 51 (2005),pp. 575-581
[9]	Chiang, S.L., Mekalanos, J.J., Holden, D.W. Annu. Rev. Microbiol., 53 (1999),pp. 129-154
[10]	Clatworthy, A.E., Pierson, E., Hung, D.T. Targeting virulence: a new paradigm for antimicrobial therapy Nat. Chem. Biol., 3 (2007),pp. 541-548
[11]	da Silva, A.C., Ferro, J.A., Reinach, F.C. et al. Comparison of the genomes of two Xanthomonas pathogens with differing host specificities Nature, 417 (2002),pp. 459-463
[12]	Daum, R.S. N. Engl. J. Med., 359 (2008),pp. 85-87
[13]	Dow, J.M., Clarke, B.R., Milligan, D.E. et al. Appl. Environ. Microbiol., 56 (1990),pp. 2994-2998
[14]	Eriksson, S., Lucchini, S., Thompson, A. et al. Mol. Microbiol., 47 (2003),pp. 103-118
[15]	He, Y.W., Boon, C., Zhou, L. et al. Mol. Microbiol., 71 (2009),pp. 1464-1476
[16]	He, Y.W., Wu, J., Zhou, L. et al. Mol. Plant Microbe Interact., 24 (2011),pp. 948-957
[17]	Innes, R.W., Hirose, M.A., Kuempel, P.L. J. Bacteriol., 170 (1988),pp. 3793-3802
[18]	Jansen, A., Yu, J. Differential gene expression of pathogens inside infected hosts Curr. Opin. Microbiol., 9 (2006),pp. 138-142
[19]	Jenkins, A., Cote, C., Twenhafel, N. et al. Infect. Immun., 79 (2011),pp. 153-166
[20]	Klarsfeld, A.D., Goossens, P.L., Cossart, P. Mol. Microbiol., 13 (1994),pp. 585-597
[21]	Krewulak, K.D., Vogel, H.J. Structural biology of bacterial iron uptake Biochim. Biophys. Acta, 1778 (2008),pp. 1781-1804
[22]	Lan, L., Cheng, A., Dunman, P.M. et al. J. Bacteriol., 192 (2010),pp. 3068-3077
[23]	Liautard, J.P., Jubier-Maurin, V., Boigegrain, R.A. et al. Antimicrobials: targeting virulence genes necessary for intracellular multiplication Trends Microbiol., 14 (2006),pp. 109-113
[24]	Liu, G.Y., Nizet, V. Color me bad: microbial pigments as virulence factors Trends Microbiol., 17 (2009),pp. 406-413
[25]	Lowe, A.M., Beattie, D.T., Deresiewicz, R.L. Mol. Microbiol., 27 (1998),pp. 967-976
[26]	Lu, G.T., Tang, Y.Q., Li, C.Y. et al. J. Bacteriol., 191 (2009),pp. 3639-3648
[27]	Mansfield, J., Genin, S., Magori, S. et al. Top 10 plant pathogenic bacteria in molecular plant pathology Mol. Plant Pathol., 13 (2012),pp. 614-629
[28]	Marra, A. Targeting virulence for antibacterial chemotherapy: identifying and characterising virulence factors for lead discovery Drugs R. D., 7 (2006),pp. 1-16
[29]	Mazurkiewicz, P., Tang, C.M., Boone, C. et al. Signature-tagged mutagenesis: barcoding mutants for genome-wide screens Nat. Rev. Genet., 7 (2006),pp. 929-939
[30]	Mei, J.M., Nourbakhsh, F., Ford, C.W. et al. Mol. Microbiol., 26 (1997),pp. 399-407
[31]	Merrell, D.S., Hava, D.L., Camilli, A. Mol. Microbiol., 43 (2002),pp. 1471-1491
[32]	Mole, B.M., Baltrus, D.A., Dangl, J.L. et al. Global virulence regulation networks in phytopathogenic bacteria Trends Microbiol., 15 (2007),pp. 363-371
[33]	Park, Y.J., Song, E.S., Kim, Y.T. et al. FEMS Microbiol. Lett., 276 (2007),pp. 55-59
[34]	Pilatz, S., Breitbach, K., Hein, N. et al. Infect. Immun., 74 (2006),pp. 3576-3586
[35]	Poplawsky, A.R., Urban, S.C., Chun, W. Appl. Environ. Microbiol., 66 (2000),pp. 5123-5127
[36]	Price, M.N., Huang, K.H., Alm, E.J. et al. A novel method for accurate operon predictions in all sequenced prokaryotes Nucleic Acids Res., 33 (2005),pp. 880-892
[37]	Qian, W., Han, Z.J., He, C. Mol. Plant Microbe Interact., 21 (2008),pp. 151-161
[38]	Qian, W., Han, Z.J., Tao, J. et al. Mol. Plant Microbe Interact., 21 (2008),pp. 1128-1138
[39]	Qian, W., Jia, Y., Ren, S.X. et al. Genome Res., 15 (2005),pp. 757-767
[40]	Rediers, H., Rainey, P.B., Vanderleyden, J. et al. Microbiol. Mol. Biol. Rev., 69 (2005),pp. 217-261
[41]	Roberts, R.L., Metz, M., Monks, D.E. et al. Proc. Natl. Acad. Sci. USA, 100 (2003),pp. 6634-6639
[42]	Ryan, R.P., Vorholter, F.J., Potnis, N. et al. Nat. Rev. Microbiol., 9 (2011),pp. 344-355
[43]	Samant, S., Lee, H., Ghassemi, M. et al. Nucleotide biosynthesis is critical for growth of bacteria in human blood PLoS Pathog., 4 (2008),p. e37
[44]	Sambrook, F.F., Maniatis, T.
[45]	Schafer, A., Tauch, A., Jager, W. et al. Gene, 145 (1994),pp. 69-73
[46]	Smith, H. Trends Microbiol., 6 (1998),pp. 239-243
[47]	Song, Y., Liu, C.I., Lin, F.Y. et al. J. Med. Chem., 52 (2009),pp. 3869-3880
[48]	Symmons, M.F., Bokma, E., Koronakis, E. et al. The assembled structure of a complete tripartite bacterial multidrug efflux pump Proc. Natl. Acad. Sci. USA, 106 (2009),pp. 7173-7178
[49]	Tang, J.L., Liu, Y.N., Barber, C.E. et al. Mol. Gen. Genet., 226 (1991),pp. 409-417
[50]	Thammavongsa, V., Kern, J.W., Missiakas, D.M. et al. J. Exp. Med., 206 (2009),pp. 2417-2427
[51]	Vorholter, F.J., Schneiker, S., Goesmann, A. et al. J. Biotechnol., 134 (2008),pp. 33-45
[52]	Wu, H.J., Wang, A.H., Jennings, M.P. Discovery of virulence factors of pathogenic bacteria Curr. Opin. Chem. Biol., 12 (2008),pp. 93-101
[53]	Wu, T., Zhao, Z., Zhang, L. et al. Microb. Pathog., 51 (2011),pp. 69-76
[54]	Zrenner, R., Stitt, M., Sonnewald, U. et al. Pyrimidine and purine biosynthesis and degradation in plants Annu. Rev. Plant Biol., 57 (2006),pp. 805-836