蛋白组学分析多聚磷酸盐在球形芽胞杆菌适应营养恢复过程中的作用

doi:10.16409/j.cnki.2095-039x.2018.03.009

中国生物防治学报 ›› 2018, Vol. 34 ›› Issue (3): 377-387.DOI: 10.16409/j.cnki.2095-039x.2018.03.009

蛋白组学分析多聚磷酸盐在球形芽胞杆菌适应营养恢复过程中的作用

石廷玉^1,3, 王园媛², 向宇波¹, 董兴高¹, 袁志明³

1. 湖北民族学院, 恩施 445000;
2. 湖北民族学院附属民大医院, 恩施 445000;
3. 中国科学院武汉病毒研究所, 武汉 430071

收稿日期:2017-07-21 出版日期:2018-06-08 发布日期:2018-06-14
通讯作者: 袁志明,博士,研究员,E-mail:yzm@wh.iov.cn
作者简介:石廷玉,博士,讲师,E-mail:shitingyu198@126.com
基金资助:
湖北民族学院博士科研启动金（MY2015B024）；湖北省教育厅科学技术研究计划青年人才项目（Q20171902）；国家自然科学基金（31600073）；湖北省大学生创新创业训练计划（201710517024）；湖北民族学院大学生创新创业训练计划项目（201810517019）；湖北省自然科学基金青年项目（2018CFB384）

Proteomics Analysis for the Role of Polyphosphate in Lysinibacillus sphaericus Adaptation to Nutrition Recovery

SHI Tingyu^1,3, WANG Yuanyuan², XIANG Yubo¹, DONG Xinggao¹, Yuan Zhiming³

1. Medical College of Hubei Institute for Nationalities, Enshi 445000, China;
2. Affiliated Hospital of Hubei Institute for Nationalities, Enshi 445000, China;
3. Key Laboratory of Agricultural and Environmental Microbiology/Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

Received:2017-07-21 Online:2018-06-08 Published:2018-06-14

摘要/Abstract

摘要： 本文通过敲除球形芽胞杆菌多聚磷酸盐合成酶基因，研究野生菌和突变菌在氨基酸缺乏以及营养恢复后细菌的生长情况。结果发现，球形芽胞杆菌野生菌在氨基酸缺乏的条件下会导致polyP聚集，而突变菌不能。在氨基酸缺乏的条件下，野生菌比突变菌生长的更好，在营养恢复后，野生菌比突变菌能更迅速地恢复生长。通过蛋白组学和生物信息学分析发现，RNA降解体组分PNP和RNase J蛋白在突变菌中上调。由于mRNA稳定性的调节是细菌适应营养环境的一种重要策略，因此，我们推测球形芽胞杆菌通过polyP的聚集，来调节mRNA稳定性，以快速适应营养缺乏和恢复后的环境。

关键词: 球形芽胞杆菌, 多聚磷酸盐, 氨基酸缺乏, 环境胁迫

Abstract: Though knocking out the gene ppk of polyphosphate kinase, the growth of Lysinibacillus sphaericus wild strains and ppk mutants in conditions of amino acids deficiency and nutrition recovery was investigated. In amino acid deficiency, L. sphaericus wild strains accumulated polyP, but L. sphaericus ppk mutants did not. The wild strains grew better than ppk mutants in amino acid deficiency medium and, the wild strains regained growth more quickly than ppk mutants in medium with nutrition recovery. Proteomics and bioinformatics analysis revealed that levels of PNP and RNase J protein, the components of RNA degradosome, were up-regulated in L. sphaericus ppk mutants. The regulation of mRNA stability was an important strategy for bacteria to adapt to nutrition deficiency, we suspect that L. sphaericus accumulates polyP to regulate mRNA stability and to adapt to nutritional deficiency and recovery environment.

Key words: Lysinibacillus sphaericus, polyphosphate, amino acids deficiency, environment stresses

中图分类号:

S476.1

石廷玉, 王园媛, 向宇波, 董兴高, 袁志明. 蛋白组学分析多聚磷酸盐在球形芽胞杆菌适应营养恢复过程中的作用[J]. 中国生物防治学报, 2018, 34(3): 377-387.

SHI Tingyu, WANG Yuanyuan, XIANG Yubo, DONG Xinggao, Yuan Zhiming. Proteomics Analysis for the Role of Polyphosphate in Lysinibacillus sphaericus Adaptation to Nutrition Recovery[J]. journal1, 2018, 34(3): 377-387.

参考文献

[1] Wu Y, Hu X, Ge Y, et al. Generation of mariner-based transposon insertion mutant library of Bacillus sphaericus 2297 and investigation of genes involved in sporulation and mosquito-larvicidal crystal protein synthesis[J]. FEMS Microbiology Letters, 2012, 330(2):105-112.
[2] Dalebroux Z D, Swanson M S. ppGpp:magic beyond RNA polymerase[J]. Microbiology, 2012, 10(3):203-212.
[3] Potrykus K, Cashel M. (p)ppGpp:still magical?[J]. Annual Review of Microbiology, 2008, 62:35-51.
[4] Vrentas C E, Gaal T, Berkmen M B, et al. Still looking for the magic spot:the crystallographically defined binding site for ppGpp on RNA polymerase is unlikely to be responsible for rRNA transcription regulation[J]. Journal of Molecular Biology, 2008, 377(2):551-564.
[5] Rao N N, Liu S, Kornberg A. Inorganic polyphosphate in Escherichia coli:the phosphate regulon and the stringent response[J]. Journal of Bacteriology, 1998, 180(8):2186-2193.
[6] Rao N N, Kornberg A. Inorganic polyphosphate supports resistance and survival of stationary-phase Escherichia coli[J]. Journal of Bacteriology, 1996, 178(5):1394-1400.
[7] Shiba T, Tsutsumi K, Yano H, et al. Inorganic polyphosphate and the induction of rpoS expression[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(21):11210-11215.
[8] Ault-Riche D, Fraley CD, Tzeng CM, et al. Novel assay reveals multiple pathways regulating stress-induced accumulations of inorganic polyphosphate in Escherichia coli[J]. Journal of Bacteriology, 1998, 180(7):1841-1847.
[9] Shi T, Ge Y, Zhao N, et al. Polyphosphate kinase of Lysinibacillus sphaericus and its effects on accumulation of polyphosphate and bacterial growth[J]. Microbiological research, 2015, 172:41-47.
[10] Neidhardt F C, Bloch P L, Smith D F. Culture medium for enterobacteria[J]. Journal of Bacteriology, 1974, 119(3):736-747.
[11] Shi X, Rao N N, Kornberg A. Inorganic polyphosphate in Bacillus cereus:motility, biofilm formation, and sporulation[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(49):17061-17065.
[12] Yuan Z, Neilsen-LeRoux C, Pasteur N, et al. Cloning and expression of the binary toxin genes of Bacillus sphaericus C3-41 in a crystal minus B. thuringiensis subsp. israelensis[J]. Acta Microbiologica Sinica, 1999, 39(1):29-35.
[13] Arantes O, Lereclus D. Construction of cloning vectors for Bacillus thuringiensis[J]. Gene, 1991, 108:115-119.
[14] Bourgouin C, Delecluse A, de la Torre F, et al. Transfer of the toxin protein genes of Bacillus sphaericus into Bacillus thuringiensis subsp. israelensis and their expression[J]. Applied and Environmental Microbiology, 1990, 56(2):340-344.
[15] Albi T, Serrano A. Inorganic polyphosphate in the microbial world. Emerging roles for a multifaceted biopolymer[J]. World Journal of Microbiology and Biotechnology, 32(2):27.
[16] Lehnik-Habrink M, Lewis R J, Mader U, et al. RNA degradation in Bacillus subtilis:an interplay of essential endo-and exoribonucleases[J]. Molecular Microbiology, 2012, 84(6):1005-1017.
[17] Schnepf E, Crickmore N, van Rie J, et al. Bacillus thuringiensis and its pesticidal crystal proteins[J]. Microbiology and Molecular Biology Reviews, 1998, 62(3):775-806.
[18] Johnson D E, McGaughey W H. Contribution of Bacillus thuringiensis spores to toxicity of purified cry proteins towards Indianmeal moth larvae[J]. Current Microbiology, 1996, 33(1):54-59.
[19] Salamitou S, Ramisse F, Brehelin M, et al. The plcR regulon is involved in the opportunistic properties of Bacillus thuringiensis and Bacillus cereus in mice and insects[J]. Microbiology, 2000, 146(11):2825-2832.
[20] Miyasono M, Inagaki S, Yamamoto M, et al. Enhancement of Delta-Endotoxin activity by toxin-free spore of Bacillus thuringiensis against the Diamondback moth, Plutella-Xylostella[J]. Journal of Invertebrate Pathology, 1994, 63(1):111-112.
[21] Donovan W P, Donovan J C, Engleman J T. Gene knockout demonstrates that vip3A contributes to the pathogenesis of Bacillus thuringiensis toward Agrotis ipsilon and Spodoptera exigua[J]. Journal of Invertebrate Pathology, 2001, 78(1):45-51.
[22] Vallet-Gely I, Lemaitre B, Boccard F. Bacterial strategies to overcome insect defences[J]. Nature Reviews Microbiology, 2008, 6(4):302-313.
[23] Azevedo C, Saiardi A. The new world of inorganic polyphosphates[J]. Biochemical Society Transactions, 2016, 44(1):13-17.
[24] Kuroda A, Murphy H, Cashel M, et al. Guanosine tetra-and pentaphosphate promote accumulation of inorganic polyphosphate in Escherichia coli[J]. The Journal of Biological Chemistry, 1997, 272(34):21240-21243.
[25] Kuroda A. A polyphosphate-lon protease complex in the adaptation of Escherichia coli to amino acid starvation Mycobacterium tuberculosis[J]. Bioscience, Biotechnology, and Biochemistry, 2006, 70(2):325-331.
[26] Sanyal S, Banerjee S K, Banerjee R, et al. Polyphosphate kinase 1, a central node in the stress response network of Mycobacterium tuberculosis, connects the two-component systems MprAB and SenX3-RegX3 and the extracytoplasmic function sigma factor, sigma E[J]. Microbiology, 2013, 159(10):2074-2086.
[27] Takayama K, Kjelleberg S. The role of RNA stability during bacterial stress responses and starvation[J]. Environmental Microbiology, 2000, 2(4):355-365.
[28] Condon C, Putzer H, Grunberg-Manago M. Processing of the leader mRNA plays a major role in the induction of thrS expression following threonine starvation in Bacillus subtilis[J]. Proceedings of the National Academy of Sciences of the United States of America, 1996, 93(14):6992-6997.
[29] Itoh H, Kawazoe Y, Shiba T. Enhancement of protein synthesis by an inorganic polyphosphate in an E. coli cell-free system[J]. Journal of Microbiological Methods, 2006, 64(2):241-249.
[30] Blum E, Py B, Carpousis A J, et al. Polyphosphate kinase is a component of the Escherichia coli RNA degradosome[J]. Molecular Microbiology, 1997, 26(2):387-398.

蛋白组学分析多聚磷酸盐在球形芽胞杆菌适应营养恢复过程中的作用

Proteomics Analysis for the Role of Polyphosphate in Lysinibacillus sphaericus Adaptation to Nutrition Recovery

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

联系我们

[1]	郭青云, 蔡全信, 胡晓敏, 闫建平, 袁志明. 球形芽胞杆菌Cry48Aa/Cry49Aa杀蚊毒素与致倦库蚊中肠上皮细胞复合物的结合特性[J]. 中国生物防治学报, 2016, 32(3): 318-325.
[2]	郭青云, 蔡全信, 胡晓敏, 闫建平, 袁志明. 球形芽胞杆菌Cry48Aa/Cry49Aa毒素在苏云金芽胞杆菌中的共表达及杀蚊活性[J]. 中国生物防治学报, 2015, 31(6): 950-955.