Journal of Guangxi Normal University(Natural Science Edition) ›› 2024, Vol. 42 ›› Issue (6): 236-243.doi: 10.16088/j.issn.1001-6600.2024022602

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Sequence Characteristics and Tissue Expression and Its Effect on Gene Expression of BCAAs Metabolic Enzymes of KLF6 Gene in Siniperca chuatsi

BIN Qin1, BIN Shiyu1,2*, BAO Lingsheng3,4, CHU Wuying3,4, ZHANG Jianshe3,4, CHENG Congyi3,4, ZENG Wei3,4, YAO Zhou3,4   

  1. 1. College of Life Sciences, Guangxi Normal University, Guilin Guangxi 541006, China;
    2. Key Laboratory of Ecology and Environmental Protection of Rare and Endangered Plants and Animals(Guangxi Normal University), Ministry of Education, Guilin Guangxi 541006, China;
    3. School of Biological and Chemical Engineering, Changsha University,Changsha Hunan 410022, China;
    4. Hunan Provincial Key Laboratory of Aquatic Animal Nutrition and Quality Control (Changsha University), Changsha Hunan 410022, China
  • Received:2024-02-26 Revised:2024-04-09 Online:2024-12-30 Published:2024-12-30

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Abstract: In order to study the sequence characteristics and tissue expression of KLF6 gene in Siniperca chuatsi and its influence on the expression of branchable amino acid metabolizing enzyme gene, bioinformatics analysis of KLF6 gene was conducted through online tools and software. Real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression abundance of KLF6 gene in 7 different tissues of the Sinfinfish and the mRNA expression levels of KLF6 gene and BCAAs metabolizing enzyme related genes after siRNA interference. The results showed that the total length of the coding region of the KLF6 gene of S. chuatsi was 3 791 bp (entry number: XM_044176561.1), in which the 5′ end and 3′ end were 271 bp and 2 659 bp, respectively, and the open reading frame was 860 bp, encoding a total of 286 amino acids. It was most closely related to the KLF6 gene (98.6%) and more distant to the KLF6 gene (82.3%) of carp. KLF6 has one KLF6-N type domain (2-204 aa), one COG5048 domain (194-268 aa) and three Znf-C2H2 domains, which were located between 203-227, 233-257 and 263-285 amino acids, respectively. The structure of the S. chuatsi KLF6 protein was composed of random curl (56.99%), α-helix (26.22%), extension chain (10.84%) and β-angle (5.94%). The results of RT-qPCR showed that the relative expression level of KLF6 gene in S. chuatsi tissues was in the following order from high to low: heart, spleen, brain, kidney, intestine, muscle and liver, and the expression level in the center was significantly higher than that in liver (P<0.01) and muscle (P<0.05), while the expression level in spleen, brain, kidney and intestine was higher than that in liver (P<0.05). The mRNA expression of BCAAs metabolizing enzymes (BCKDHb, ACADM, MCCC1, MCCC2, MUT, IVD) was significantly increased after the mRNA expression of KLF6 gene was inhibited (P<0.05). This study provides a basis for the study of the regulatory mechanism of KLF6 gene in muscle BCAAs metabolism of S. chuatsi.

Key words: Siniperca chuatsi, transcription factor, branched-chain amino acid, metabolic enzyme, tissue expression

CLC Number:  S917.4
[1] SYAFRUDDIN S E, MOHTAR M A, NAZARIE W F W M, et al. Two sides of the same coin: the roles of KLF6 in physiology and pathophysiology[J]. Biomolecules, 2020, 10(10): 1378. DOI: 10.3390/biom10101378.
[2] HSIEH P N, FAN L Y, SWEET D R, et al. The Krüppel-like factors and control of energy homeostasis[J]. Endocrine Reviews, 2019,40(1):137-152. DOI: 10.1210/er.2018-00151.
[3] D’ASTOLFO D S, GEHRAU R C, BOCCO J L, et al. Silencing of the transcription factor KLF6 by siRNA leads to cell cycle arrest and sensitizes cells to apoptosis induced by DNA damage[J]. Cell Death & Differentiation, 2008, 15(3): 613-616. DOI: 10.1038/sj.cdd.4402299.
[4] 庄兆辉, 仲永, 陈月婵, 等. Krüppel样因子在肌肉组织中的功能研究进展[J]. 遗传, 2018, 40(9): 733-748. DOI: 10.16288/j.yczz.18-095.
[5] 陈知龙. 猪KLF家族基因克隆及分子特性研究[D]. 武汉: 华中农业大学, 2010.
[6] YU L, ZHENG W J, LI C Q, et al. Mir-22-3p/KLF6/MMP14 axis in fibro-adipogenic progenitors regulates fatty infiltration in muscle degeneration[J]. The FASEB Journal, 2020, 34(9): 12691-12701. DOI: 10.1096/fj.202000506R.
[7] 康亭亭. 猪骨骼肌卫星细胞增殖分化过程中的基因表达调控网络及KLF6基因的功能研究[D]. 武汉: 华中农业大学, 2020.
[8] SONG C C, FANG X, YANG Z X, et al. MiR-152 regulates bovine myoblast proliferation by targeting KLF6[J]. Animals, 2021, 11(10): 3001. DOI: 10.3390/ani11103001.
[9] 陈妮, 赵梅. MiR-181a-5p通过靶向KLF6信号调控低氧诱导的肺动脉平滑肌细胞的增殖和迁移的机制研究[J]. 系统医学, 2022, 7(24): 30-36, 58. DOI: 10.19368/j.cnki.2096-1782.2022.24.030.
[10] PIRET S E, GUO Y Q, ATTALLAH A A, et al. Krüppel-like factor 6-mediated loss of BCAA catabolism contributes to kidney injury in mice and humans[J]. Proceedings of the National Academy of Sciences of the United States of America, 2021, 118(23): e2024414118. DOI: 10.1073/pnas.2024414118.
[11] 朱静华, 丛林. 浅谈支链氨基酸的功效和使用[J]. 田径, 2022(5): 84, 75.
[12] BROSNAN J T, BROSNAN M E.Branched-chain amino acids: enzyme and substrate regulation[J]. The Journal of Nutrition, 2006, 136(1): 207S-211S. DOI: 10.1093/jn/136.1.207S.
[13] 王开卓, 宾石玉, 李虹辉, 等. 翘嘴鳜生长抑制素基因的克隆及其表达分析[J]. 广西师范大学学报(自然科学版), 2013, 31(4):109-114. DOI: 10.16088/j.issn.1001-6600.2013.04.017.
[14] 朱鑫, 叶苏杭, 李源, 等. 鳜miR-21的时空表达特征及短期饥饿胁迫下的适应性节律表达调控[J]. 中国水产科学, 2022, 29(5): 665-672. DOI: 10.12264/JFSC2021-0520.
[15] SUGO T, TERADA M, OIKAWA T, et al. Development of antibody-siRNA conjugate targeted to cardiac and skeletal muscles[J]. Journal of Controlled Release, 2016, 237: 1-13. DOI: 10.1016/j.jconrel.2016.06.036.
[16] 胡姜丽, 徐蕾, 许艺兰, 等. 鳜Follistatin基因cDNA的克隆及其胚胎发育表达分析[J]. 广西师范大学学报(自然科学版), 2020, 38(4): 124-131. DOI: 10.16088/j.issn.1001-6600.2020.04.015.
[17] 王江林, 王永, 孟庆勇, 等. 山羊KLF6的分子克隆及其时空表达特征分析[J]. 华北农学报, 2021, 36(2): 226-232. DOI: 10.7668/hbnxb.20191543.
[18] BLACK A R, BLACK J D,AZIZKHAN-CLIFFORD J. Sp1 and Krüppel-like factor family of transcription factors in cell growth regulation and cancer[J]. Journal of Cellular Physiology, 2001, 188(2): 143-160. DOI: 10.1002/jcp.1111.
[19] 周佳, 孙志鹏, 吉宇丹, 等. 梭鲈ghrh基因序列特征及其在大小个体间差异表达分析[J]. 淡水渔业, 2023, 53(5): 68-76. DOI: 10.3969/j.issn.1000-6907.2023.05.008.
[20] LAUB F, ALDABE R, RAMIREZ F, et al. Embryonic expression of Krüppel-like factor 6 in neural and non-neural tissues[J]. Mechanisms of Development, 2001, 106(1/2): 167-170. DOI: 10.1016/s0925-4773(01)00419-1.
[21] 贾鲁, 薛良义, 蔡灿, 等. 大黄鱼KLF6基因分子克隆和特征分析[J]. 生物学杂志, 2012, 29(1): 5-8. DOI: 10.3969/j.issn.2095-1736.2012.01.005.
[22] 张锐, 唐纪全. 胃癌组织中KLF6和PTTG1的表达与胃癌淋巴结转移及预后的关系[J]. 中国现代普通外科进展, 2023, 26(11): 869-872, 885. DOI: 10.3969/j.issn.1009-9905.2023.11.007.
[23] GHIASSI-NEJAD Z, HERNANDEZ-GEA V, WOODRELL C, et al. Reduced hepatic stellate cell expression of Krüppel-like factor 6 tumor suppressor isoforms amplifies fibrosis during acute and chronic rodent liver injury[J]. Hepatology, 2013, 57(2): 786-796. DOI: 10.1002/hep.26056.
[24] SAWAKI D, HOU L G, TOMIDA S, et al. Modulation of cardiac fibrosis by Krüppel-like factor 6 through transcriptional control of thrombospondin 4 in cardiomyocytes[J]. Cardiovascular Research, 2015, 107(4): 420-430. DOI: 10.1093/cvr/cvv155.
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