DrwH类信号肽序列对其抗氧化功能的影响

doi:10.13560/j.cnki.biotech.bull.1985.2018-1050

生物技术通报 ›› 2019, Vol. 35 ›› Issue (5): 125-132.doi: 10.13560/j.cnki.biotech.bull.1985.2018-1050

DrwH类信号肽序列对其抗氧化功能的影响

郭磊周¹, 韩佳慧², 唐殷¹, 李江^1,2, 黄程¹, 代其林¹, 王劲², 平淑珍², 江世杰¹

1.西南科技大学生命科学与工程学院,绵阳 621010;
2.中国农业科学院生物技术研究所,北京 100081

收稿日期:2018-12-10 出版日期:2019-05-26 发布日期:2019-05-23
作者简介:郭磊周,男,硕士研究生,研究方向：特殊环境微生物功能基因资源利用;E-mail：13548463016@163.com
基金资助:
国家自然科学基金项目（31800061）,西南科技大学博士基金（18zx7127）

Effect of Signal Peptide-like Sequence in DrwH on Its Antioxidant Function

GUO Lei-zhou¹, HAN Jia-hui², TANG Yin¹, LI Jiang^1,2, HUANG Cheng¹, DAI Qi-lin¹, WANG Jin², PING Shu-zhen², JIANG Shi-jie¹

1.College of Life Science and Engineering,Southwest University of Science and Technology,Mianyang 621010;
2.Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081

Received:2018-12-10 Published:2019-05-26 Online:2019-05-23

摘要/Abstract

摘要： 耐辐射异常球菌疏水性LEA5C家族蛋白DrwH参与细胞耐受氧化和盐胁迫过程。前期研究发现,DrwH蛋白N端存在19个氨基酸组成的类信号肽序列,为探索该序列对DrwH发挥抗氧化功能的影响,本研究通过对DrwH蛋白进行分段截短,构建含有不同区段的重组大肠杆菌菌株,比较分析重组菌株在正常培养和氧化胁迫下的生长能力。结果表明,含有类信号肽序列的融合蛋白过量表达不仅抑制了大肠杆菌的生长,而且使DrwH蛋白失去抗氧化能力。荧光定量PCR显示经IPTG诱导后,含类信号肽全长蛋白DrwH和结构域蛋白WHy在转录水平大量累积,蛋白在过量积累条件下发挥其生物学功能。由此表明,类信号肽对全长蛋白DrwH表达及其发挥抗氧化功能具有重要影响。

关键词: 耐辐射异常球菌, DrwH蛋白, WHy结构域, 类信号肽, 抗氧化

Abstract: DrwH,a hydrophobic LEA5C protein of Deinococcus radiodurans,is involved in the process of cell tolerance to oxidative and salinity stresses. Previous studies suggest the presence of a signal peptide-like sequence of 19 amino acids in the N-terminal of DrwH protein. In order to explore the effect of the signal peptide-like sequence on the antioxidant function of DrwH,DrwH protein was truncated to different fragments and then recombinant Escherichia coli strains containing the different fragments were constructed,and their growths were analyzed and compared between normal culture and oxidative stress. The results showed that overexpression of the fusion protein containing the signal peptide-like sequence not only inhibited the growth of E. coli,but also caused DrwH protein to lose its antioxidant capacity. qRT-PCR results demonstrated that,after induction by IPTG,the full-length protein DrwH containing signal peptide-like sequence and the WHy-domain were accumulated at the transcriptional level,and the proteins exerted its biological function under excessive accumulation. This indicates that the signal peptide-like sequence presents an important influence on the expression of the full-length protein DrwH and its antioxidant function.

Key words: Deinococcus radiodurans, DrwH protein, WHy domain, signal peptide-like, antioxidant function

郭磊周, 韩佳慧, 唐殷, 李江, 黄程, 代其林, 王劲, 平淑珍, 江世杰. DrwH类信号肽序列对其抗氧化功能的影响[J]. 生物技术通报, 2019, 35(5): 125-132.

GUO Lei-zhou, HAN Jia-hui, TANG Yin, LI Jiang, HUANG Cheng, DAI Qi-lin, WANG Jin, PING Shu-zhen, JIANG Shi-jie. Effect of Signal Peptide-like Sequence in DrwH on Its Antioxidant Function[J]. Biotechnology Bulletin, 2019, 35(5): 125-132.

参考文献

[1] Makarova KS, Aravind L, Wolf YI, et al.Genome of the extremely radiation-resistant bacterium Deinococcus radiodurans viewed from the perspective of comparative genomics[J]. Microbiology and Molecular Biology Reviews, 2001, 65(1):44-79.
[2] Mattimore V, Battista JR.Radioresistance of Deinococcus radiodurans:functions necessary to survive ionizing radiation are also necessary to survive prolonged desiccation[J]. Journal of Bacteriology, 1996, 178(3):633-637.
[3] Lewis NF.Studies on a Radio-resistant Coccus isolated from Bombay duck(Harpodon nehereus)[J]. Microbiol, 1971, 1:29-35.
[4] Omelchenko MV, Wolf YI, Gaidamakova EK, et al.Comparative genomics of Thermus thermophilus and Deinococcus radiodurans:divergent routes of adaptation to thermophily and radiation resistance[J]. BMC Evolutionary Biology, 2005, 5(1):57.
[5] Jiang SJ, et al.DrwH, a novel WHy domain-containing hydrophobic LEA5C protein from Deinococcus radiodurans, protects enzymatic activity under oxidative stress[J]. Sci Rep, 2017, 1:9281.
[6] Sharma A, et al.Ectopic expression of an atypical hydrophobic group 5 LEA protein from wild peanut, Arachis diogoi confers abiotic stress tolerance in tobacco[J]. PLoS One, 2016, 11(3):e0150609.
[7] He S, Tan L, Hu Z, et al.Molecular characterization and functional analysis by heterologous expression in E. coli under diverse abiotic stresses for OsLEA5, the atypical hydrophobic LEA protein from Oryza sativa L.[J]. Mol Genet Genomics, 2012, 287(1):39-54.
[8] 张亨, 刘盈盈, 陈云, 等. 戈壁异常球菌 Dgl5 蛋白生物学功能研究[J]. 生物技术通报, 2018, 34(3):177-184.
[9] Ciccarelli FD, Bork P.The Why domain mediates the response to desiccation in plants and bacteria[J]. Bioinformatics, 2004, 21(8):1304-1307.
[10] Anderson D, Ferreras E, et al. A novel bacterial Water Hypersensi-tivity-like protein shows in vivo protection against cold and freeze damage[J]. FEMS Microbiol Lett, 2015, 362(15):fnv110.
[11] Liang Y, Xiong Z, Zheng J, et al.Genome-wide identification, structural analysis and new insights into late embryogenesis abundant(LEA)gene family formation pattern in Brassica napus[J]. Scientific Reports, 2016, 6:24265.
[12] Ling H, Zeng X, Guo S.Functional insights into the late embryogenesis abundant(LEA)protein family from Dendrobium officinale(Orchidaceae)using an Escherichia coli system[J]. Scientific Reports, 2016, 6:39693.
[13] Liu Y, Zheng Y.PM2, a group 3 LEA protein from soybean, and its 22-mer repeating region confer salt tolerance in Escherichia coli[J]. Biochem Biophys Res Commun, 2005, 1:325-332.
[14] Zou Y, Hong R, He S, et al.Polyproline II structure is critical for the enzyme protective function of soybean Em(LEA1)conserved domains[J]. Biotechnology Letters, 2011, 33(8):1667-1673.
[15] 徐妙云, 刘德虎, 李刚强. 大豆24 kDa油体蛋白基因在大肠杆菌中的表达[J]. 中国农业科学, 2005(2):234-240.
[16] Nguyen HQ, Le PD, Chu HM.High-level expression, purification and properties of an Endochitinase gene without signal peptide from Lecanicillium lecanii 43H in Pichia pastoris[J]. Molecular Biology Reports, 2018, 45(5):1067-1075.
[17] Karlsson M, et al.A sequence-specific function for the N-terminal signal-like sequence of the TonB protein[J]. Mol Microbiol, 1993, 2:379-388.
[18] 彭蔚宇, 等. 去除信号肽有利鸡白细胞介素-10在哺乳动物细胞胞浆的表达[J]. 实验动物科学, 2017, 34:16-20.
[19] Liu MX, Allegood J, Zhu XW, et al.Uncleaved apoM signal peptide is required for formation of large apoM/S1P-enriched HDL particles[J]. J Biol Chem, 2015, 290(12):7861-7870.
[20] Wang Y, Zheng J, Lin FL, et al.Improved extracellular endo-1, 4-β-mannosidase activity of recombinant Pichia pastoris by optimizing signal peptide[J]. J Cent South Univ, 2015, 22(6):2088-2095.

DrwH类信号肽序列对其抗氧化功能的影响

Effect of Signal Peptide-like Sequence in DrwH on Its Antioxidant Function

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