Direct influence of the p53 tumor suppressor on mitochondrial biogenesis and function
Direct influence of the p53 tumor suppressor on mitochondrial biogenesis and function
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DOI：
10.1096/fj.00-0262com
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发表时间：
2001-03-01
期刊：
影响因子：
4.8
通讯作者：
Knudsen, TB
中科院分区：
文献类型：
--
作者：
Donahue, RJ;Razmara, M;Knudsen, TB
在一些细胞系统中已经观察到p53的线粒体定位，但对其基于细胞器的生理活性的理解仍然不完整。本研究的目的是研究线粒体DNA基因组对融合到线粒体输入信号的显性阴性p53突变体微小蛋白（p53DD）的反应。通过与小鼠cox81的信号肽序列进行框内融合，生成表达线粒体靶向增强型绿色荧光蛋白（mEGFP）或显性阴性突变型p53微小蛋白（m53DD）的构建体。对照胞质载体（cEGFP、c53DD）的信号序列位于反义方向。将这些载体以不同组合瞬时转染NIH 3T3细胞。在表达m53DD的细胞中，线粒体16S核糖体RNA （16S rRNA）的表达和Mitotracker Red CMXRos （Delta Psim）的荧光染色均降低，这两种改变对线粒体导入能力（例如，m53DD vs. c53DD）以及乘客蛋白（例如，m53DD vs. mEGFP）都是特异性的。线粒体外周型苯二氮卓受体的特异性配体PK11195恢复了线粒体的正常功能状态。m53DD对16S rRNA表达和CMXRos染色的负显性，以及PK11195对这些参数的挽救，表明p53对线粒体的生物发生和功能有直接的积极作用。-Donahue， R. J, Razmara， M., Hock， J. B, Knudsen， T. B.肿瘤抑制因子p53对线粒体生物发生和功能的直接影响。
Mitochondrial localization of p53 has been observed in several cell systems, but an understanding of its organelle-based physiological activity remains incomplete. The purpose of the present study was to investigate the mitochondrial DNA genomic response to dominant-negative p53 mutant miniprotein (p53DD) fused to a mitochondrial import signal. Constructs were generated to express mitochondrial targeted enhanced green fluorescent protein (mEGFP) or dominant-negative mutant p53 miniprotein (m53DD) by in-frame fusion to the signal peptide sequence of murine Cox8l. Control cytosolic vectors (cEGFP, c53DD) had the signal sequence placed in antisense orientation. NIH 3T3 cells were transiently transfected with these vectors in various combinations. Mitochondrial 16S ribosomal RNA (16S rRNA) expression and fluorochrome staining with Mitotracker Red CMXRos (Delta Psim) were decreased in cells expressing m53DD, Both alterations were specific for mitochondrial import competence (e.g., m53DD vs. c53DD) as well as the passenger protein (e.g., m53DD vs. mEGFP). The normal functional state of mitochondria was restored with PK11195, a specific ligand of the mitochondrial peripheral-type benzodiazepine receptor. Negative dominance of m53DD on 16S rRNA expression and CMXRos staining, and rescue of these parameters with PK11195, imply a direct positive effect of p53 on mitochondrial biogenesis and function.-Donahue, R. J., Razmara, M., Hock, J. B., Knudsen, T. B. Direct influence of the p53 tumor suppressor on mitochondrial biogenesis and function.