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| The small and large subunits of carbamoyl-phosphate synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri |
| Guo Jing, SonG Xue, Zou Li-fang, Zou Hua-song, CHen Gong-you |
| School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P.R.China |
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摘要 Carbamoyl-phosphate synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the small and large subunits of carbamoyl-phosphate synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hypersensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracellular enzyme activities. These data demonstrated that only the large subunit of carbamoyl-phosphate synthase was essential for canker development by X. citri subsp. citri.
Abstract Carbamoyl-phosphate synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the small and large subunits of carbamoyl-phosphate synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hypersensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracellular enzyme activities. These data demonstrated that only the large subunit of carbamoyl-phosphate synthase was essential for canker development by X. citri subsp. citri.
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Received: 21 July 2014
Accepted:
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| Fund: This work was supported by the National Natural Science Foundation of China (31171832) and the Special Fund for Agro-Scientific Research in the Public Interest, China (201003067). |
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Corresponding Authors:
ZOU Hua-song, Tel: +86-21-34205873,E-mail: hszou@sjtu.edu.cn; CHEN Gong-you, Tel: +86-21-34205873, E-mail: gyouchen@sjtu.edu.cn
E-mail: hszou@sjtu.edu.cn;gyouchen@sjtu.edu.cn
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Cite this article:
Guo Jing, SonG Xue, Zou Li-fang, Zou Hua-song, CHen Gong-you.
2015.
The small and large subunits of carbamoyl-phosphate synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri. Journal of Integrative Agriculture, 14(7): 1338-1347.
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| Ahuja V, Powers-Lee S G. 2008. Human carbamoyl-phosphatesynthetase: Insight into N-acetylglutamate interactionand the functional effects of a common single nucleotidepolymorphism. Journal of Inherited Metabolic Disease, 31,481-491Alegria M C, Docena C, Khater L, Ramos C H, da Silva AC R, Farah C S. 2004. New protein-protein interactionsidentified for the regulatory and structural componentsand substrates of the type III secretion system of thephytopathogen Xanthomonas axonopodis pathovar citri.Journal of Bacteriology, 186, 6186-6197AI-Saadi A, Reddy J D, Duan Y P, Brunings A M, YuanQ, Gabriel D W. 2007. All five host-range variants ofXanthomonas citri carry one pthA homolog with 17.5 repeatsthat determines pathogenicity on citrus, but none determinehost-range variation. Molecular Plant-Microbe Interactions,20, 934-943Arioli S, Monnet C, Guglielmetti S, Mora D. 2009.Carbamoylphosphate synthetase activity is essential forthe optimal growth of Streptococcus thermophilus in milk.Journal of Applied Microbiology, 107, 348-354Beckwith J R, Pardee A B, Austrian R, Jacob F. 1962.Coordination of the synthesis of the enzymes in thepyrimidine pathway of E. coli. Journal of Molecular Biology,5, 618-634Bouvier J, Patte J C, Stragier P. 1984. Multiple regulatorysignals in the control region of the Escherichia coli carABoperon. Proceedings of the National Academy of Sciencesof the United States of America, 81, 4139-4143Charlier D, Hassanzadeh G, Kholti A, Gigot D, Pierard A,Glansdorff N. 1995. carP, involved in pyrimidine regulationof the Escherichia coli carbamoylphosphate synthetaseoperon, encodes a sequence-specific DNA-binding proteinidentical to XerB and PepA, also required for resolutionof ColEI multimers. Journal of Molecular Biology, 250,392-406Charlier D, Roovers M, Gigot D, Huysveld N, Pierard A,Glansdorff N. 1993. Integration host factor (IHF) modulatesthe expression of the pyrimidine-specific promoter of thecarAB operons of Escherichia coli K-12 and Salmonellatyphimurium LT2. Molecular and General Genetics, 237,273-286Crouser E D, Julian M W, Huff J E, Struck J, Cook CH. 2006. Carbamoyl phosphate synthase-1:marker of mitochondrial damage and depletion inthe liver during sepsis. Critical Care Medicine, 34, 2439-2546Devroede N, Thia-Toong T L, Gigot D, Maes D, Charlier D. 2004.Purine and pyrimidine-specific repression of the Escherichiacoli carAB operon are functionally and structurally coupled.Journal of Molecular Biology, 336, 25-42Dzierszinshi F S, Hunter C A. 2008. Advances in the use ofgenetically engineered parasites to study immunity toToxoplasma gondii. Parasite Immunology, 30, 235-244Dunger G, Arabolaza A L, Gottig N, Orellano E G, Ottado J.2005. Participation of Xanthomonas axonopodis pv. citri hrpcluster in citrus canker and nonhost plant responses. PlantPathology, 54, 781-788Dunger G, Garofalo C G, Gottig N, Garavaglia B S, Rosa M CP, Farah C S, Orellano E G, Ottado J. 2012. Analysis ofthree Xanthomonas axonopodis pv. citri effector proteins inpathogenicity and their interactions with host plant proteins.Molecular Plant Pathology, 13, 865-876Guo Y P, Figueiredo F, Jones J, Wang N. 2011. HrpG and HrpXplay global roles in coordinating different virulence traits ofXanthomonas axonopodis pv. citri. Molecular Plant-MicrobeInteractions, 24, 649-661Han S, Auger C, Appanna V P, Lemire J, CastonguayZ, Akbarov E, Appanna V D. 2012. A blue nativepolyacrylamide gel electrophoretic technology to probethe functional proteomics mediating nitrogen homeostasisin Pseudomonas fluorescens. Journal of MicrobiologicalMethods, 90, 206-210Holden H M, Thoden J B, Raushel F M. 1998. Carbamoylphosphate synthetase: a tunnel runs through it. CurrentOpinion in Structural Biology, 8, 679-685Holden H M, Thoden J B, Raushel F M. 1999. Carbamoylphosphate synthetase: An amazing biochemical odysseyfrom substrate to product. Cellular and Molecular LifeSciences, 56, 507-522Hu Y, Zhang J, Jia H, Sosso D, Li T, Frommer W B, YangB, White F F, Wang N, Jones J B. 2014. Lateral organboundaries 1 is a disease susceptibility gene for citrusbacterial canker disease. Proceedings of the NationalAcademy of Sciences of the United States of America,111, E521-E529.Jones M E, Spector L. 1960. The pathway of carbonate in thebiosynthesis of carbamyl phosphate. Journal of BiologicalInorganic Chemistry, 235, 2897-2901Kholti A, Charlier D, Gigot D, Huysveld N, Roovers M,Glansdorff N. 1998. pyrH-encoded UMP-kinase directlyparticipates in pyrimidine-specific modulation of promoteractivity in Escherichia coli. Journal of Molecular Biology,280, 571-582Kim J, Howell S, Huang X, Raushel F M. 2002. Structuraldefects within the carbamate tunnel of carbamoyl phosphatesynthetase. Biochemistry, 41, 12575-12581Koo B S, Hyun H H, Kim S Y, Kim C H, Lee H C. 2011.Enhancement of thymidine production in E. coli byeliminating repressors regulating the carbamoyl phosphatesynthetase operon. Biotechnology Letters, 33, 71-78Kovach M E, Phillips R W, Elzer P H, Roop R M, Peterson KM. 1994. pBBR1MCS: A broad-host-range cloning vector.Biotechniques, 16, 800-802Li J Y, Wang N. 2011. Genome-wide mutagenesis ofXanthomonas axonopodis pv. citri reveals novel geneticdeterminants and regulation mechanisms of biofilmformation. PLoS One, 6, e21804.Malamud F, Homem R A, Conforte V P, Yaryura P M,Castagnaro A P, Marano M R, do Amaral A M, VojnovA A. 2013. Identification and characterization of biofilmformation-defective mutants of Xanthomonas citri subsp.citri. Microbiology, 159, 1911-1919McGivan J D, Bradford N M, Mendes-Mourão J. 1976. Theregulation of carbamoyl phosphate synthase activity in ratliver mitochondria. Biochemical Journal, 154, 415-421Ono H, Suto T, Kinoshita Y, Sakano T, Furue T, Ohta T. 2009.A case of carbamoyl phosphate synthetase I deficiencypresenting symptoms at one month of age. Brain &Development, 31, 779-781Raushel F M, Thoden J B, Holden H M. 1999. Theamidotransferase family of enzymes: Molecular machinesfor the production and delivery of ammonia. Biochemistry,38, 7891-7899Rigano L A, Siciliano F, Enrique R, Sendín L, Filippone P,Torres P S, Qüesta J, Dow J M, Castagnaro A P, Vojnov AA, Marano M R. 2007. Biofilm formation, epiphytic fitness,and canker development in Xanthomonas axonopodis pv.citri. Molecular Plant-Microbe Interactions, 20, 1222-1230Peng L, Hertz L, Huang R, Sonnewald U, Petersen S B,Westergaard N, Larsson O, Schousboe A. 1993. Utilizationof glutamine and of TCA cycle constituents as precursorsfor transmitter glutamate and GABA. DevelopmentalNeuroscience, 15, 367-377Piette J, Nyunoya H, Lusty C J, Cunin R, Weyens G, CrabeelM. 1984. DNA sequence of the carA gene and the controlregion of carAB: tandem promoters, respectively controlledby arginine and the pyrimidines, regulate the synthesis ofcarbamoyl-phosphate synthetase in Escherichia coli K-12Proceedings of the National Academy of Sciences of theUnited States of America, 81, 4134-4138Schulte R, Bonas U 1992. The expression of the hrp genecluster from Xanthomonas campestris pv. vesicatoria thatdetermines pathogenicity and hypersensitivity on pepperand tomato is plant-inducible. Journal of Bacteriology,174, 815-823Sgro G G, Ficarra F A, Dunger G, Scarpec T E, Valle EM, Cortadi A, Orellano E G, Gottig N, Ottado J2012.Contribution of a harpin protein from Xanthomonasaxonopodis pv. citri to pathogen virulence. Molecular PlantPathology, 13, 1047-1059Shimada T, Hirao K, Kori A, Yamamoto K, Ishihama A2007.RutR is the uracil/thymine-sensing master regulator of aset of genes for synthesis and degradation of pyrimidines.Molecular Microbiology, 66, 744-757Siciliano F, Torres P, Send?n L, Bermejo C, Filippone P, VelliceG, Ramalo J, Castagnaro A, Vojnov A, Marano M R 2006.Analysis of the molecular basis of Xanthomonas axonopodispv. citri pathogenesis in citrus limon. Electronic Journal ofBiotechnology, 9, 200-204Stapleton M A, Javid-Majd F, Harmon M F, Hanks B A,Grahmann J L, Mullins L S, Raushel F M 1996. Role ofconserved residues within the carboxy phosphate domainof carbamoyl phosphate synthetase. Biochemistry, 35,14352-14361Thoden J B, Raushel F M, Benning M M, Tayment I, Holden H M 1999. The structure of carbamoyl phosphate synthetasedetermined to 2.1 A resolution. Acta Crystallographica(Section D, Biological Crystallography), 55, 8-24Tondo M, Petrocelli S, Ottado J, Orellano E. 2010. Themonofunctional catalase KatE of Xanthomonas axonopodispv. citri is required for full virulence in citrus plants. PLoSOne, 5, e10803.Vauterin L, Hoste B, Kersters K, Swings J. 1995. Reclassificationof Xanthomonas. International Journal of SystematicBacteriology, 45, 472-489Wengelnik K, Bonas U. 1996. HrpXv, an AraC-type regulator,activates expression of five of the six loci in the hrp clusterof Xanthomonas campestris pv. vesicatoria. Journal ofBacteriology, 178, 3462-3469Yan Q, Wang N. 2012. High-throughput screening and analysisof genes of Xanthomonas citri subsp. citri involved in citruscanker symptom development. Molecular Plant-MicrobeInteractions, 25, 69-84Zimaro T, Thomas L, Marondedze C, GaravagliaB S, Gehring C, Ottado J, Gottig N. 2013. Insightsinto Xanthomonas axonopodis pv. citri biofilm throughproteomics. BMC Microbiology, 13, 186.Zou H S, Song X, Zou L F, Yuan L, Li Y R, Guo W, CheY Z, Zhao W X, Chen G Y, Duan Y P. 2012. EcpA, anextracellular protease, is a specific virulence factor requiredby Xanthomonas oryzae pv. oryzicola but not by X. oryzaepv. oryzae in rice. Microbiology, 158, 2372-2383 |
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