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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林乃君(Nai-Chun Lin) | |
dc.contributor.author | Li Kang Sung | en |
dc.contributor.author | 宋立綱 | zh_TW |
dc.date.accessioned | 2021-06-17T04:27:36Z | - |
dc.date.available | 2019-08-18 | |
dc.date.copyright | 2018-08-18 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-13 | |
dc.identifier.citation | 1. Polz MF, Alm EJ, & Hanage WP (2013) Horizontal gene transfer and the evolution of bacterial and archaeal population structure. Trends Genet 29:170-175.
2. Baltrus DA (2013) Exploring the costs of horizontal gene transfer. Trends Ecol Evol 28:489-495. 3. Will WR, Navarre WW, & Fang FC (2015) Integrated circuits: how transcriptional silencing and counter-silencing facilitate bacterial evolution. Curr Opin Microbiol 23:8-13. 4. Gordon BR, Li YF, Cote A, Weirauch MT, Ding PF, Hughes TR, Navarre WW, Xia B, & Liu J (2011) Structural basis for recognition of AT-rich DNA by unrelated xenogeneic silencing proteins. Proc Natl Acad Sci U S A 108:10690-10695. 5. Jacquet M, Cukier-Kahn R, Pla J, & Gros F (1971) A thermostable protein factor acting on in vitro DNA transcription. Biochem Biophys Res Commun 45:1597-1607. 6. Varshavsky A, Nedospasov S, Bakayev V, Bakayeva T, & Georgiev G (1977) Histone-like proteins in the purified Escherichia coli deoxyribonucleoprotein. Nucleic Acids Res 4:2725-2746. 7. Goyard S & Bertin P (1997) Characterization of BpH3, an H-NS-like protein in Bordetella pertussis. Mol Microbiol 24:815-823. 8. Ussery W, Hinton D, Jorji M, Granum E, Seirafi A, Stephen J, Tupper A, Berridge G, Sidebotham JM, & Higgins CF (1994) The chromatin-associated protein H-NS. Biochimie 76:968-980. 9. Tendeng C, Badaut C, Krin E, Gounon P , Ngo S , Danchin A , Rimsky S, & Bertin P (2000) Isolation and characterization of vicH, encoding a new pleiotropic regulator in Vibrio cholerae. J Bacteriol 182:2026-2032. 10. Müller CM, Dobrindt U, Nagy G, Emödy L, Uhlin BE, & Hacker J (2006) Role of histone-like proteins H-NS and StpA in expression of virulence determinants of uropathogenic Escherichia coli. Journal of bacteriology 188(J Bacteriol):5428-5438. 11. Bertin P, Terao E, Lee EH, Lejeune P, Colson C, Danchin A , & Collatz E (1994) The H-NS protein is involved in the biogenesis of flagella in Escherichia coli. J Bacteriol 176:5537-5540. 12. Laurent-Winter C, Ngo S, Danchin A, & Bertin P (1997) Role of Escherichia coli histone-like nucleoid-structuring protein in bacterial metabolism and stress response: identification of targets by two-dimensional electrophoresis. Eur J Biochem 244:767-773. 13. Navarre WW, Porwollik S, Wang YP, McClell M, Rosen H, Libby SJ, & Fang FC (2006) Selective silencing of foreign DNA with low GC content by the H-NS protein in Salmonella. Science 313:236-238. 14. Wilmes-Riesenberg MR, Foster JW, & Curtiss R (1997) An altered rpoS allele contributes to the avirulence of Salmonella typhimurium LT2. Infect Immun 65:203-210. 15. Lucchini S, Rowley G, Goldberg MD, Hurd D, Harrison M, & Hinton JC (2006) H-NS mediates the silencing of laterally acquired genes in bacteria. PLoS Pathog 2:e81. 16. Tendeng C, Soutourina O, Danchin A, & Bertin P (2003) MvaT proteins in Pseudomonas spp.: a novel class of H-NS-like proteins. Microbiology 149:3047-3050. 17. Ding PF, McFarland KA, Jin SJ, Tong G, Duan B, Yang A, Hughes TR, Liu J, Dove CL, Navarre WW, & Xia B (2015) A novel AT-rich DNA recognition mechanism for bacterial xenogeneic silencer MvaT. PLoS Pathog 11:e1004967. 18. Wiggins PA, Dame RT, Noom MC, & Wuite GJ (2009) Protein-mediated molecular bridging: a key mechanism in biopolymer organization. Biophys J 97:1997-2003. 19. Diggle SP, Winzer K, Lazdunski A, Williams P, & Cámara M (2002) Advancing the quorum in Pseudomonas aeruginosa: MvaT and the regulation of N-acylhomoserine lactone production and virulence gene expression. J Bacteriol 184:2576-2586. 20. Vallet I, Diggle SP, Stacey RE, Cámara M, Ventre I, Lory S, Lazdunski A, Williams P, & Filloux A (2004) Biofilm formation in Pseudomonas aeruginosa: fimbrial cup gene clusters are controlled by the transcriptional regulator MvaT. J bacteriol 186:2880-2890. 21. Vallet-Gely I, Donovan KE, Fang R, Joung JK, & Dove SL (2005) Repression of phase-variable cup gene expression by H-NS-like proteins in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 102:11082-11087. 22. Castang S, McManus HR, Turner KH, & Dove SL (2008) H-NS family members function coordinately in an opportunistic pathogen. Proc Natl Acad Sci U S A 105:18947-18952. 23. Li C, Wally H, Miller SJ, & Lu C-D (2009) The multifaceted proteins MvaT and MvaU, members of the H-NS family, control arginine metabolism, pyocyanin synthesis, and prophage activation in Pseudomonas aeruginosa PAO1. J Bacteriol 191:6211-6218. 24. Renzi F, Rescalli E, Galli E, & Bertoni G (2010) Identification of genes regulated by the MvaT-like paralogues TurA and TurB of Pseudomonas putida KT2440. Environ Microbiol 12:254-263. 25. Baehler E, Werra P, Wick LY, Péchy-Tarr M, Mathys S, Maurhofer M, & Keel C (2006) Two novel MvaT-like global regulators control exoproduct formation and biocontrol activity in root-associated Pseudomonas fluorescens CHA0. Mol Plant Microbe Interact 19:313-329. 26. Gardan L, Shafik H, Belouin S, Broch R, Grimont F, & Grimont PA (1999) DNA relatedness among the pathovars of Pseudomonas syringae and description of Pseudomonas tremae sp. nov. and Pseudomonas cannabina sp. nov. (ex Sutic and Dowson 1959). Int J Syst Evol Microbiol 49:469-478. 27. Dangl JL & Jones JD (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826. 28. Whalen MC, Innes RW, Bent AF, & Staskawicz BJ (1991) Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell 3:49-59. 29. Bronstein PA, Marrichi M, Cartinhour S, Schneider DJ, & DeLisa MP (2005) Identification of a twin-arginine translocation system in Pseudomonas syringae pv. tomato DC3000 and its contribution to pathogenicity and fitness. Journal Bacteriol 187:8450-8461. 30. Schulze-Lefert P & Robatzek S (2006) Plant pathogens trick guard cells into opening the gates. Cell 126:831-834. 31. Alfano JR, Charkowski AO, Deng WL, Badel JL, Petnicki-Ocwieja T, Dijk K, & Collmer A (2000) The Pseudomonas syringae Hrp pathogenicity island has a tripartite mosaic structure composed of a cluster of type III secretion genes bounded by exchangeable effector and conserved effector loci that contribute to parasitic fitness and pathogenicity in plants. Proc Natl Acad Sci U S A 97:4856-4861. 32. Sarris PF, Skandalis N, Kokkinidis M, & Panopoulos NJ (2010) In silico analysis reveals multiple putative type VI secretion systems and effector proteins in Pseudomonas syringae pathovars. Mol Plant Pathol 11:795-804. 33. Haapalainen M, Mosorin H, Dorati F, Wu RF, Roine E, Taira S, Nissinen R, Mattinen L, Jackson R, Pirhonen M, & Lin NC (2012) Hcp2, a secreted protein of the phytopathogen Pseudomonas syringae pv. tomato DC3000, is required for fitness for competition against bacteria and yeasts. J Bacteriol 194:4810-4822. 34. Maffei B, Francetic O, & Subtil A (2017) Tracking Proteins Secreted by Bacteria: What's in the Toolbox? Front Cell Infect Microbiol 7:221. 35. Green ER & Mecsas J (2016) Bacterial secretion systems–an overview. Microbiol Spectr 4. 36. Cornelis GR (2006) The type III secretion injectisome. Nat Reviews Microbiol 4:811. 37. Diepold A & Armitage JP (2015) Type III secretion systems: the bacterial flagellum and the injectisome. Phil. Trans. R. Soc. B 370:20150020. 38. Alfano JR & Collmer A (2004) Type III secretion system effector proteins: double agents in bacterial disease and plant defense. Annu. Rev. Phytopathol. 42:385-414. 39. Xin X-F, Kvitko B, & He SY (2018) Pseudomonas syringae: what it takes to be a pathogen. Nat Revi Microbiol. 40. Stauber JL, Loginicheva E, & Schechter LM (2012) Carbon source and cell density-dependent regulation of type III secretion system gene expression in Pseudomonas syringae pathovar tomato DC3000. Res Microbiol 163:531-539. 41. Hutcheson SW, Bretz J, Sussan T, Jin S, & Pak K (2001) Enhancer-binding proteins HrpR and HrpS interact to regulate hrp-encoded Type III protein secretion in Pseudomonas syringae strains. J Bacteriology 183:5589-5598. 42. Fouts DE, Abramovitch RB, Alfano JR, Baldo AM, Buell CR, Cartinhour S, Chatterjee AK, D'Ascenzo M, Gwinn ML, Lazarowitz SG, Lin NC, Martin GB, Rehm AH, Schneider DJ, Dijk K, Tang X, & Collmer A (2002) Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor. Proc Natl Acad Sci U S A 99:2275-2280. 43. Lan L, Deng X, Zhou J, & Tang X (2006) Genome-wide gene expression analysis of Pseudomonas syringae pv. tomato DC3000 reveals overlapping and distinct pathways regulated by hrpL and hrpRS. Mol Plant Microbe Interact 19:976-987. 44. Ronald PC, Salmeron J, Carland FM, & Staskawicz B (1992) The cloned avirulence gene avrPto induces disease resistance in tomato cultivars containing the Pto resistance gene. J Bacteriol 174:1604-1611. 45. Martin G, de Vicente MC, & Tanksley S (1993) High-resolution linkage analysis and physical characterization of the Pto bacterial resistance locus in tomato. Mol Plant Microbe Interact. 46. Xiang T, Zong N, Zhang Z, Chen J, Chen M, & Zhou JM (2011) BAK1 is not a target of the Pseudomonas syringae effector AvrPto. Mol Plant Microbe Interact 24:100-107. 47. Xiang T, Zong N, Wu Y, Zhang J, Xing W, Li Y, Tang X, Zhu L, Chai J, & Zhou JM (2008) Pseudomonas syringae effector AvrPto blocks innate immunity by targeting receptor kinases. Curr Biol 18:74-80. 48. Hauck P, Thilmony R, & He SY (2003) A Pseudomonas syringae type III effector suppresses cell wall-based extracellular defense in susceptible Arabidopsis plants. Proc Natl Acad Sci U S A 100:8577-8582. 49. Bingle LE, Bailey CM, & Pallen MJ (2008) Type VI secretion: a beginner's guide. Curr Opin Microbiol 11:3-8. 50. Pukatzki, Ma AT, Sturtevant D, Krastins B, Sarracino D, Nelson WC, Heidelberg JF, & Mekalanos JJ (2006) Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proceedings of the National Academy of Sciences 103(Proc Natl Acad Sci U S A):1528-1533. 51. Cascales E (2008) The type VI secretion toolkit. EMBO Rep 9:735-741. 52. Jani AJ & Cotter PA (2010) Type VI secretion: not just for pathogenesis anymore. Cell Host Microbe 8:2-6. 53. Chen H, Yang D, Han F, Tan J, Zhang L, Xiao J, Zhang Y, & Liu Q (2017) The bacterial T6SS effector EvpP prevents NLRP3 inflammasome activation by inhibiting the Ca2+-dependent MAPK-Jnk pathway. Cell Host Microbe 21:47-58. 54. Si M, Zhao C, Burkinshaw B, Zhang B, Wei D, Wang Y, Dong TG, & Shen X (2017) Manganese scavenging and oxidative stress response mediated by type VI secretion system in Burkholderia thailandensis. Proc Natl Acad Sci U S A 114:2233-2242. 55. Wang T, Si M, Song Y, Zhu W, Gao F, Wang Y, Zhang L, Zhang W, Wei G, Luo ZQ, & Shen X (2015) Type VI secretion system transports Zn2+ to combat multiple stresses and host immunity. PLoS Pathog 11:e1005020. 56. Lin J, Zhang W, Cheng J, Yang X, Zhu K, Wang Y, Wei G, Qian PY, Luo ZQ, & Shen X (2017) A Pseudomonas T6SS effector recruits PQS-containing outer membrane vesicles for iron acquisition. Nat Commun:14888. 57. Mougous JD, Cuff ME, Raunser S, Shen A, Zhou M, Gifford CA, Goodman AL, Joachimiak G, Ordoñez CL, Lory S, Walz T, Joachimiak A, & Mekalanos JJ (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science 312:1526-1530. 58. Williams S, Varcoe L, Attridge SR, & Manning PA (1996) Vibrio cholerae Hcp, a secreted protein coregulated with HlyA. Infect Immun 64:283-289. 59. Basler á, Pilhofer á, Henderson G, Jensen G, & Mekalanos J (2012) Type VI secretion requires a dynamic contractile phage tail-like structure. Nature 483:182. 60. Pukatzki S, Ma AT, Revel AT, Sturtevant D, & Mekalanos JJ (2007) Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc Natl Acad Sci U S A 104:15508-15513. 61. Leiman PG, Basler M, Ramagopal UA, Bonanno LB, Sauder JM, Pukatzki S, Burley SK, Almo SC, & Mekalanos JJ (2009) Type VI secretion apparatus and phage tail-associated protein complexes share a common evolutionary origin. Proc Natl Acad Sci U S A 106:4154-4159. 62. Cianfanelli FR, Monlezun L, & Coulthurst SJ (2016) Aim, load, fire: the type VI secretion system, a bacterial nanoweapon. Trends Microbiol 24:51-62. 63. Suarez G, Sierra JC, Erova TE, Sha J, Horneman AJ, & Chopra AK (2010) A type VI secretion system effector protein, VgrG1, from Aeromonas hydrophila that induces host cell toxicity by ADP ribosylation of actin. J Bacteriol 192:155-168. 64. Schwarz S, Singh P, Robertson JD, LeRoux M, Skerrett3 SJ, Goodlett DR, West TE, & Mougous JD (2014) VgrG-5 is a Burkholderia type VI secretion exported protein required for multinucleated giant cell formation and virulence. Infect Immun:IAI. 01368-01313. 65. Russell AB, LeRoux M, Hathazi K, Agnello DM, Ishikawa T, Wiggins PA, Wai SN, & Mougous JD (2013) Diverse type VI secretion phospholipases are functionally plastic antibacterial effectors. Nature 496:508-512. 66. Russell AB, Hood RD, Bui NK, LeRoux M, Vollmer W, & Mougous JD (2011) Type VI secretion delivers bacteriolytic effectors to target cells. Nature 475:343-347. 67. Alcoforado DJ & Coulthurst S (2015) Intraspecies Competition in Serratia marcescens Is Mediated by Type VI-Secreted Rhs Effectors and a Conserved Effector-Associated Accessory Protein. J Bacteriol 197:2350-2360. 68. Basler á & Mekalanos J (2012) Type VI secretion dynamics within and between bacterial cells. Science 337:815-815. 69. Cianfanelli FR, Diniz JA, Guo M, Cesare V, Trost M, & Coulthurst SJ (2016) VgrG and PAAR proteins define distinct versions of a functional type VI secretion system. PLoS Pathog 12:e1005735. 70. Liang X, Moore R, Wilton M, Wong MJ, Lam L, & Dong TG (2015) Identification of divergent type VI secretion effectors using a conserved chaperone domain. Proc Natl Acad Sci U S A 112:9106-9111. 71. Ma J, Pan Z, Huang J, Sun M, Lu C, &Yao H (2017) The Hcp proteins fused with diverse extended-toxin domains represent a novel pattern of antibacterial effectors in type VI secretion systems. Virulence 8:1189-1202. 72. Brooks TM, Unterweger D, Bachmann V, Kostiuk B, & Pukatzki S (2013) Lytic activity of the Vibrio cholerae type VI secretion toxin VgrG-3 is inhibited by the antitoxin TsaB. J Biol Chem 288:7618-7625. 73. Russell AB, Peterson SB, & Mougous JD (2014) Type VI secretion system effectors: poisons with a purpose. Nat Rev Microbiol 12:137. 74. Bondage DD, Lin JS, Ma LS, Kuo CH, & Lai EM (2016) VgrG C terminus confers the type VI effector transport specificity and is required for binding with PAAR and adaptor–effector complex. Proc Natl Acad Sci U S A:201600428. 75. English G, Trunk K, Rao VA, Srikannathasan V, Hunter WN, & Coulthurst SJ (2012) New secreted toxins and immunity proteins encoded within the Type VI secretion system gene cluster of Serratia marcescens. Mol Microbiol 86:921-936. 76. Ma LS, Hachani A, Lin JS, Filloux A, & Lai EM (2014) Agrobacterium tumefaciens deploys a superfamily of type VI secretion DNase effectors as weapons for interbacterial competition in planta. Cell Host Microbe 16:94-104. 77. Lien YW & Lai EM (2017) Type VI secretion effectors: Methodologies and biology. Front Cell Infect Microbiol 7:254. 78. Chien CF (2017) Characterization of the T6SS HSI-II gene cluster in Pseudomonas syringae pv. tomato DC3000. Doctor thesis. 79. Chen KY (2015) Characterization of the type VI secretion system VgrG proteins in Pseudomonas syringae pv. tomato DC3000. Master thesis. 80. Lu YY (2015) Characterization of PSPTO_0281 gene in phytogenic Pseudomonas syringae pv. tomato DC3000. Master thesis. 81. Santana FJ, Calva E, & Puente JL (2001) Transcriptional regulation of type III secretion genes in enteropathogenic Escherichia coli: Ler antagonizes H-NS-dependent repression. Mol Microbiol 39:664-678. 82. Troxell B, Sikes ML, Fink RC1, Vazquez-Torres A, Jones-Carson J, & Hassan HM (2011) Fur negatively regulates hns and is required for the expression of HilA and virulence in Salmonella enterica serovar Typhimurium. J Bacteriol 193:497-505. 83. Zhang J, Xiao J, Zhang Y, Cui S, Liu Q, Wang Q, Wu H, & Zhang Y (2014) A new target for the old regulator: H-NS suppress T6SS secretory protein EvpP, the major virulence factor in the fish pathogen Edwardsiella tarda. Lett Appl Microbiol 59:557-564. 84. Sana TG, Flaugnatti N, Lugo KA, Lam LH, Jacobson A, Baylot V, Durand E, Journet L, Cascales E, & Monack DM (2016) Salmonella Typhimurium utilizes a T6SS-mediated antibacterial weapon to establish in the host gut. Proc Natl Acad Sci U S A 113:E5044-E5051. 85. Kim J, Lee JY, Lee H, Choi JY, Kim DH, Wi YM, Peck KR, & Ko KS (2017) Microbiological features and clinical impact of the type VI secretion system (T6SS) in Acinetobacter baumannii isolates causing bacteremia. Virulence 8:1378-1389. 86. Salomon D, Klimko JA, & Orth K (2014) H-NS regulates the Vibrio parahaemolyticus type VI secretion system 1. Microbiology 160:1867-1873. 87. Hsu CH, Chen C, Jou ML, Lee YL, Lin YC, Yu YP, Huang WT, & Wu SH (2005) Structural and DNA-binding studies on the bovine antimicrobial peptide, indolicidin: evidence for multiple conformations involved in binding to membranes and DNA. Nucleic Acids Res 33:4053-4064. 88. Lin NC & Martin GB (2005) An avrPto/avrPtoB mutant of Pseudomonas syringae pv. tomato DC3000 does not elicit Pto-mediated resistance and is less virulent on tomato. Mol Plant Microbe Interact 18:43-51. 89. Saitou N & Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406-425. 90. Kelley LA, Mezulis S, Yates CM, Wass MN, & Sternberg MJ (2015) The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc 10:845. 91. Suzuki-Minakuchi C, Kawazuma K, Matsuzawa J, Vasileva D, Fujimoto Z, Terada T, Okada K, & Nojiri H (2016) Structural similarities and differences in H-NS family proteins revealed by the N-terminal structure of TurB in Pseudomonas putida KT2440. FEBS Lett 590:3583-3594. 92. Lee YC (2011) Development and application of a new site-specific chromosomal integration strategy for gene expression in Pseudomonas syringae pv. tomato DC3000. Master thesis. 93. Juhas M, Meer JR, Gaillard M, Harding RM, Hood DW, & Crook DW (2009) Genomic islands: tools of bacterial horizontal gene transfer and evolution. FEMS Microbiol Rev 33:376-393. 94. Winsor GL, Griffiths EJ, Lo R, Dhillon BK, Shay JA, & Brinkman FS (2015) Enhanced annotations and features for comparing thousands of Pseudomonas genomes in the Pseudomonas genome database. Nucleic Acids Res 44:D646-D653. 95. Cui S, Xiao J, Wang Q, & Zhang Y (2016) H-NS binding to evpB and evpC and repressing T6SS expression in fish pathogen Edwardsiella piscicida. Arch Microbiol 198:653-661. 96. Gallagher SR (2012) GUS protocols: using the GUS gene as a reporter of gene expression (Academic Press). 97. Dorman CJ (2004) H-NS: a universal regulator for a dynamic genome. Nat Rev Microbiol 2:391. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70411 | - |
dc.description.abstract | 水平基因轉移 (Horizontal gene transfer, HGT) 係指遺傳物質除了從親代傳至子代之外,於生物間移動的現象,為演化動力之一,可使生物獲取新性狀,以適應環境。然而,若未經適當調控,而一味表現這些外來基因,會給予宿主壓力,並可能導致自身適性下降。而為了因應外源基因可能造成的負面效應,細菌演化出一特別之轉錄抑制子,稱為外源基因沉默子 (xenogeneic silencer),可結合上潛在之外源基因序列,以調控其表現。此外源基因沉默子可按其序列分成三大類:分別是 Proteobacteria、Mycobacteria 及 Pseudomonas 屬所特有的轉錄抑制子 H-NS、Lsr2 及 MvaT。番茄細菌性斑點病菌 Pseudomonas syringae pv. tomato (Pst) DC3000 是一桿狀、具鞭毛的革蘭氏陰性菌,可在自然環境下感染番茄,其致病力依賴細菌第三型分泌系統,而與其他細菌競爭時則有第六型分泌系統參與其中。第三型分泌系統組成蛋白的基因群座落於一典型、由三部分所組成的病原性小島 (pathogenicity island) 之 hrp/hrc 基因叢集中;HSI-I 和 HSI-II 基因叢集則負責編碼第六型分泌系統的組成蛋白,於該二分泌系統之基因叢集內均可發現轉位酶 (transposase) 的基因,暗示兩叢集有可能是經由水平基因轉移獲得的外來序列。本研究在 Pst DC3000 中找到 PSPTO_0281、PSPTO_3103、PSPTO_4315 和 PSPTO_4755 四個 MvaT 之同源基因,經突變株建構與細菌間競爭試驗發現,PSPTO_3103 會負向調控 Pst DC3000 之細菌間競爭能力。後經由半定量反轉錄聚合酶連鎖反應、GUS 活性測試及西方墨點法,發現 PSPTO_3103 可抑制第六型分泌系統基因叢集及第三型分泌系統相關基因之表現,並影響胞內 Hcp2 之生產。此外,PSPTO_3103 突變株的致病力較野生株更高,說明其對於致病力的影響。透過凝膠遷移試驗,亦證明 PSPTO_3103 可與第六型基因叢集中三個操作子、hrpL及 avrPto 之啟動子結合。總而言之, PSPTO_3103 應為 Pst DC3000 中重要之 MvaT 同源蛋白,負責調控其可能為外來序列之細菌第三型及第六型分泌系統基因表現,進而影響 Pst DC3000對番茄的致病性,以及和其他細菌間的競爭能力。 | zh_TW |
dc.description.abstract | Horizontal gene transfer (HGT) is a movement of genetic materials between organisms except transmission of DNA from parents to offspring, and could serve as a driving force for evolution. HGT is thought to be used to acquire new phenotypes for bacteria to adapt to the changing environments. However, inappropriate regulation of foreign genes obtained by HGT may lead to reduction of bacterial fitness. To avoid potential problems caused by foreign DNA, bacteria have evolved the so-called “xenogeneic silencers” to regulate the expressions of those foreign genes. The xenogeneic silencers can be divided into three groups based on their sequences, which are histone-like nucleoid structuring (H-NS) protein in Proteobacteria, Lsr2 in mycobacteria, and MvaT-like protein in Pseudomonas spp. Pseudomonas syringae pv. tomato (Pst) DC3000 is a rod-shaped, Gram-negative bacterium with polar flagella, and a phytopathogen that can cause tomato speck disease. In Pst DC3000, ability to infect host plants requires its type III secretion system (T3SS) while competing with other bacteria depends on full functions of type VI secretion system (T6SS). T3SS is encoded by hrp (hypersensitive response and pathogenicity) and hrc (hrp and conserved) genes within the canonical tripartite pathogenicity islands (T-PAIs), and core components of T6SS are encoded by genes in Hcp secretion island (HSI)-I and HSI-II gene clusters. It is noteworthy that genes for transposases are within or flanked the gene clusters of hrp/hrc, HSI-I, and HSI-II, suggesting that they might be acquired by HGT, and could be putative targets of xenogeneic silencers. In this study, four MvaT orthologs in Pst DC3000, namely PSPTO_0281, PSPTO_3103, PSPTO_4315, and PSPTO_4755, were identified. By means of mutagenesis and an interbacterial competition assay, we revealed that PSPTO_3103 negatively controls the competition ability of Pst DC3000. Results from semiquantitative RT-PCR, the GUS promoter assay and Western blotting analysis demonstrated that PSPTO_3103 represses the expressions of HSI-II gene clusters, hrpL and avrPto as well as Hcp2 production. Deletion of PSPTO_3103 also increases the virulence of Pst DC3000. Furthermore, the gel shift assay indicated that PSPTO_3103 can bind to the promoter regions of operons in HSI-II gene cluster, hrpL and avrPto in a non-specific manner. Taking together, we conclude that PSPTO_3103 is the key MvaT ortholog responsible for modulating the expressions of putative foreign sequences, i.e. T3SS and T6SS-related genes, in Pst DC3000, further controlling its virulence and interbacterial competition ability. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T04:27:36Z (GMT). No. of bitstreams: 1 ntu-107-R05623010-1.pdf: 2423983 bytes, checksum: 711befaa5d7fd43980364502352a0ece (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | Index
Abstract I 中文摘要 III Introduction 1 Histone-like nucleoid structuring (H-NS) protein 1 MvaT 3 Pseudomonas syringae pv. tomato DC3000 5 Type III secretion system in Pst DC3000 6 Type VI secretion system in Pst DC3000 8 Purpose of this study 11 Materials and methods 13 Bacteria strains, media, and growth conditions 13 Growth condition of tomato plants 13 Genomic DNA isolation 14 Construction of recombinant plasmids for in-frame deletion/complementation of PSPTO_3103 15 RNA extraction 16 Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) 17 Interbacterial competition assay 18 GUS activity assay 19 Expression and purification of C-terminal His6-tagged PSPTO_3103 20 Gel shift assay 21 Protein analysis 22 Pathogenicity assay 23 Statistical analysis 24 Results 25 Four MvaT orthologs present in Pst DC3000 resemble MvaT or MvaU 25 PSPTO_0281, PSPTO_3103, and PSPTO_4315 are constitutively expressed in Pst DC3000 26 PSPTO_3103 negatively controls the interbacterial competition ability 27 PSPTO_3103 contains a conserved oligomerization domain and a DNA binding domain of MvaT family 28 PSPTO_3103 represses the expression of all operons in the HSI-II gene clusters 30 Deletion of PSPTO_3103 increases the virulence of Pst DC3000 31 PSPTO_3103 represses the gene expression of hrpL and avrPto 32 PSPTO_3103 binds to the promoter regions with a non-specific manner 33 Chromosomal integragion of C-terminal FLAG-tagged PSPTO_3103 functions as the native protein partially 35 Discussion 37 References 44 List of tables and figures Tables Table 1. Bacterial strains used in this study 60 Table 2. Primers used in this study 61 Table 3. Plasmids used in this study 62 Figures Figure 1. Multiple amino acid sequence alignment of MvaT orthologs in Pseudomonas spp. 63 Figure 2. The Neighbor-joining phylogenetic tree of MvaT orthologs in Pseudomonas spp.. 64 Figure 3. The expression levels of MvaT orthologs in Pseudomonas syringae pv. tomato DC3000 detected by semi-quantitative RT-PCR 65 Figure 4. Numbers of Escherichia coli MG1655 recovered in the interbacterial competition assay with different genotypes of Pseudomonas syringae pv. tomato DC3000. E. coli MG1655 was used as the target cells to test the interbacterial competition ability of Pst DC3000 66 Figure 5. The predicted protein structure of PSPTO_3103 67 Figure 6. Reduction in interbacterial competition ability of ∆3103 can be restored in a complementary strain 68 Figure 7. PSPTO_3103 negative regulates the expression of three operons in Hcp secretion island (HSI)-II 69 Figure 8. Changes in the expression levels of three operons in HSI-II of ∆3103 can be restored in a complementary strain 70 Figure 9. The protein level of Hcp2 increases in ∆3103 71 Figure 10. PSPTO_3103 negatively regulates virulence of Pseudomonas syringae pv. tomato DC3000 in tomato cultivar Money Maker 72 Figure 11. The expressions of hrpL and avrPto were not affected by PSPTO_3103 in a GUS reporter assay 73 Figure 12. The expression levels of hrpL, avrPto, and hcp2 in different genotypes of Pseudomonas syringae pv. tomato DC3000 74 Figure 13. Purification of C-terminal His6-tagged PSPTO_3103 by Ni-NTA affinity chromatography 75 Figure 14. PSPTO_3103 binds to the promoter regions with a non-specific manner. 76 Figure 15. Reduction in competition ability of ∆3103 can be restored by C-terminal FLAG-tagged PSPTO_3103. C-terminal FLAG-tagged PSPTO_3103 was constructed for ChIP-seq analysis 77 | |
dc.language.iso | zh-TW | |
dc.title | 植物病原細菌 Pseudomonas syringae pv. tomato DC3000
中 MvaT 同源蛋白 PSPTO_3103 之特性分析 | zh_TW |
dc.title | Characterization of PSPTO_3103, an MvaT-like protein, in phytopathogen Pseudomonas syringae pv. tomato DC3000 | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄧文玲(Wen?Ling Deng),吳蕙芬(Whei-Fen Wu),鄭秋萍(Chiu-Ping Cheng) | |
dc.subject.keyword | Pseudomonas syringae,第三型分泌系統,第六型分泌系統,MvaT-like protein,外源基因沉默子, | zh_TW |
dc.subject.keyword | Pseudomonas syringae,type III secretion system (T3SS),type VI secretion system (T6SS),MvaT-like protein,xenogeneic silencer, | en |
dc.relation.page | 77 | |
dc.identifier.doi | 10.6342/NTU201802190 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-14 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農業化學研究所 | zh_TW |
顯示於系所單位: | 農業化學系 |
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