[1]赵 兰,黄 鹰.粟酒裂殖酵母中Mmp1蛋白的功能研究[J].南京师范大学学报(工程技术版),2022,22(04):069-74.[doi:10.3969/j.issn.1672-1292.2022.04.009]
　Zhao Lan,Huang Ying.Studies on the Function of Mmp1 Protein in Schizosaccharomyces Pombe[J].Journal of Nanjing Normal University(Engineering and Technology),2022,22(04):069-74.[doi:10.3969/j.issn.1672-1292.2022.04.009]
点击复制

粟酒裂殖酵母中Mmp1蛋白的功能研究
分享到：

南京师范大学学报（工程技术版）[ISSN:1006-6977/CN:61-1281/TN]

卷:
22卷
期数:
2022年04期
页码:
069-74
栏目:
生物工程
出版日期:
2022-12-15

文章信息/Info

Title:
Studies on the Function of Mmp1 Protein in Schizosaccharomyces Pombe
文章编号:
1672-1292(2022)04-0069-06
作者:
赵 兰; 黄 鹰
(南京师范大学生命科学学院,江苏 南京 210023)
Author(s):
Zhao Lan; Huang Ying
(School of Life Sciences,Nanjing Normal University,Nanjing 210023,China)
关键词:
Mmp1; 线粒体; 实时荧光定量PCR; 线粒体蛋白提取
Keywords:
Mmp1; mitochondria; qRT-PCR; mitochondrial protein extraction
分类号:
Q78
DOI:
10.3969/j.issn.1672-1292.2022.04.009
文献标志码:
A
摘要:
线粒体是存在于大多数真核细胞中的一种产能细胞器,它参与了细胞内ATP的合成、物质的代谢、细胞凋亡等多种生理过程. Mmp1蛋白被预测为线粒体内膜肽酶复合体催化亚基,但其具体功能尚未得到解析. 通过分子克隆、表型实验、荧光显微观察、线粒体分离提取、实时荧光定量PCR、蛋白质印迹法等技术,分析了Mmp1的定位及其对线粒体功能的影响. 通过荧光定位分析发现Mmp1是一个定位在线粒体中的蛋白. 细胞缺失mmp1无法在非发酵型培养基上生长,暗示缺失mmp1影响了线粒体的功能. 同时,细胞缺失mmp1,使线粒体基因组编码的呼吸链复合体核心蛋白Cox1、Cox2、Cox3、Cob1、Atp6的蛋白水平显著降低. 在对野生型和Δmmp1菌株线粒体编码的基因的mRNA水平进行实时荧光定量实验发现,Δmmp1菌株中的Cox1、Cox2、Cox3、Cob1、Atp6的转录水平并未有明显变化. 结果表明,Mmp1影响了线粒体呼吸链复合体亚基的蛋白水平,对维持线粒体功能的发挥起着重要作用.
Abstract:
Mitochondria are the productive organelles of most eukaryotic cells,which are involved in various physiological processes such as ATP synthesis,substance metabolism and cell apoptosis. Mmp1 protein is predicted to be the catalytic subunit of the mitochondrial endopeptidase complex,but its specific function has not been analyzed. In this paper,the localization of Mmp1 and its effect on mitochondrial function are analyzed by molecular cloning,phenotypic experiment,fluorescence microscopic observation,mitochondrial isolation and extraction,real-time fluorescence quantitative PCR and western-blot. Fluorescence localization analysis reveal that Mmp1 is a protein located in mitochondria. Δmmp1 strain can not grow on the medium with glycerol as carbon source and can not perform aerobic respiration,suggesting that the deletion of mmp1 affects mitochondrial function. Meanwhile,the protein levels of Cox1,Cox2,Cox3,Cob1 and Atp6,the core proteins of respiratory chain complex encoded by mitochondrial genome,are significantly reduced when mmp1 is knock out. The transcriptional levels of Cox1,Cox2,Cox3,Cob1 and Atp6 in Δmmp1 strain do not change significantly. These results indicate that Mmp1 affects the protein levels of some mitochondrial respiratory chain complex subunits and plays an important role in maintaining mitochondrial function.

参考文献/References:

[1]KWON Y Y,CHOI K M,CHO C Y,et al. Mitochondrial efficiency-dependent viability of Saccharomyces cerevisiae mutants carrying individual electron transport chain component deletions[J]. Mol Cells,2015,38(12):1054-1063.
[2]NESCI S,PAGLIARANI A,ALGIERI C,et al. Mitochondrial F-type ATP synthase:multiple enzyme functions revealed by the membrane-embedded FO structure[J]. Critical Reviews Biochemistry and Molecular Biology,2020,55(4):309-321.
[3]COYNE L P,CHEN X J. Consequences of inner mitochondrial membrane protein misfolding[J]. Mitochondrion,2019,49:46-55.
[4]PAGLIARINI D J,RUTTER J. Hallmarks of a new era in mitochondrial biochemistry[J]. Genes & Development,2013,27(24):2615-2627.
[5]SPINELLI J B,HAIGIS M C. The multifaceted contributions of mitochondria to cellular metabolism[J]. Nature Cell Biology,2018,20(7):745-754.
[6]HUANG M L H,CHIANG S,KALINOWSKI D S,et al. The role of the antioxidant response in mitochondrial dysfunction in degenerative diseases:cross-talk between antioxidant defense,autophagy,and apoptosis[J]. Oxid Med Cell Longev,2019,2019:6392763.
[7]CHAN D C. Mitochondria:dynamic organelles in disease,aging,and development[J]. Cell,2006,125(7):1241-1252.
[8]JEFFARES D C. The natural diversity and ecology of fission yeast[J]. Yeast,2018,35(3):253-260.
[9]STEFANATOS R,SANZ A. The role of mitochondrial ROS in the aging brain[J]. FEBS Letter,2018,592(5):743-758.
[10]ANNESLEY S J,FISHER P R. Mitochondria in health and disease[J]. Cells,2019,8(7):680.
[11]SOUSA J S,D'IMPRIMA E,VONCK J. Mitochondrial respiratory chain complexes[J]. Subcellular Biochemistry,2018,87:167-227.
[12]NOLFI-DONEGAN D,BRAGANZA A,SHIVA S. Mitochondrial electron transport chain:Oxidative phosphorylation,Oxidant production,and methods of measurement[J]. Redox Biology,2020,37:101674.
[13]YAN C J,DUANMU X Y,ZENG L,et al. Mitochondrial DNA:distribution,mutations,and elimination[J]. Cells,2019,8(4):379.
[14]HERBERT C J,LABARRE-MARIOTTE S,CORNU D,et al. Translational activators and mitoribosomal isoforms cooperate to mediate mRNA-specific translation in Schizosaccharomyces pombe mitochondria[J]. Nucleic Acids Research,2021,49(19):11145-11166.
[15]LIU J Y,LI Y,CHEN J,et al. The fission yeast Schizosaccharomyces pombe Mtf2 is required for mitochondrial Cox1 gene expression[J]. Microbiology,2018,164(3):400-409.
[16]ANZIANO P Q,PERLMAN P S,LANG B F,et al. The mitochondrial genome of the fission yeast schizosaccharomyces pombe:I. isolation and physical mapping of mitochondrial DNA[J]. Current Genetics,1983,7(4):273-84.
[17]BULLERWELL C E,LEIGH J,FORGET L,et al. A comparison of three fission yeast mitochondrial genomes[J]. Nucleic Acids Research,2003,31(2):759-68.
[18]SHANG J J,YANG Y M,WU L,et al. The S. pombe mitochondrial transcriptome[J]. RNA,2018,24(9):1241-1254.
[19]JAJOO R,JUNG Y,HUH D,et al. Accurate concentration control of mitochondria and nucleoids[J]. Science,2016,351(6269):169-172.
[20]MONTANARI A,BESAGNI C,DE LUCA C,et al. Yeast as a model of human mitochondrial tRNA base substitutions:investigation of the molecular basis of respiratory defects[J]. RNA,2008,14(2):275-283.
[21]LUO Y,WANG Y R,HUANG Y. Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation[J]. Journal of Biological Chemistry,2021,297(1):100869.

相似文献/References:

[1]张 云,黄 鹰.粟酒裂殖酵母Atp25蛋白定位和功能研究[J].南京师范大学学报(工程技术版),2021,21(04):080.[doi:10.3969/j.issn.1672-1292.2021.04.012]
　Zhang Yun,Huang Ying.Study on the Localization and Function of Atp25 Proteinin Schizosaccharomyces Pombe[J].Journal of Nanjing Normal University(Engineering and Technology),2021,21(04):080.[doi:10.3969/j.issn.1672-1292.2021.04.012]
[2]王子璇,商巾杰.粟酒裂殖酵母中Hsp60蛋白定位与表达水平的研究[J].南京师范大学学报(工程技术版),2021,21(04):087.[doi:10.3969/j.issn.1672-1292.2021.04.013]
　Wang Zixuan,Shang Jinjie.Localization and Expression of Hsp60 Protein in Schizosaccharomyces pombe[J].Journal of Nanjing Normal University(Engineering and Technology),2021,21(04):087.[doi:10.3969/j.issn.1672-1292.2021.04.013]

备注/Memo

备注/Memo:
收稿日期:2022-02-28.
基金项目:国家自然科学基金项目(31400012).
通讯作者:黄鹰,博士,教授,研究方向:微生物学. E-mail:hyhanghai1965@163.com
更新日期/Last Update: 2022-12-15