Mayall T P, Bjarnason I, Khoo U Y, Peters T J, Macpherson A J
Department of Clinical Biochemistry, King's College School of Medicine and Dentistry, London, U.K.
Biochem J. 1995 Jun 1;308 ( Pt 2)(Pt 2):665-71. doi: 10.1042/bj3080665.
Most mitochondrial genes are transcribed as a single large transcript from the heavy strand of mitochondrial DNA, and are subsequently processed into the proximal mitochondrial (mt) 12 S and 16 S rRNAs, and the more distal tRNAs and mRNAs. We have shown that in intestinal epithelial biopsies the steady-state levels of mt 12 S and 16 S rRNA are an order of magnitude greater than those of mt mRNAs. Fractionation of rat small intestinal epithelial cells on the basis of their maturity has shown that the greatest ratios of 12 S mt rRNA/cytochrome b mt mRNA or 12 S mt rRNA/cytochrome oxidase I mt mRNA are found in the surface mature enterocytes, with a progressive decrease towards the crypt immature enteroblasts. Cytochrome b and cytochrome oxidase I mt mRNA levels are relatively uniform along the crypt-villus axis, but fractionation experiments showed increased levels in the crypt base. The levels of human mitochondrial transcription factor A are also greater in immature crypt enteroblasts compared with mature villus enterocytes. These results show that the relative levels of mt rRNA and mRNA are distinctly regulated in intestinal epithelial cells according to the crypt-villus position and differentiation status of the cells, and that there are higher mt mRNA and mt TFA levels in the crypts, consistent with increased transcriptional activity during mitochondrial biogenesis in the immature enteroblasts.
大多数线粒体基因作为一个单一的大转录本从线粒体DNA的重链转录而来,随后被加工成近端线粒体(mt)12S和16S rRNA,以及更远端的tRNA和mRNA。我们已经表明,在肠道上皮活检中,mt 12S和16S rRNA的稳态水平比mt mRNA的稳态水平高一个数量级。根据大鼠小肠上皮细胞的成熟度进行分级分离显示,在表面成熟的肠上皮细胞中,12S mt rRNA/细胞色素b mt mRNA或12S mt rRNA/细胞色素氧化酶I mt mRNA的比例最高,向隐窝未成熟的成肠细胞逐渐降低。细胞色素b和细胞色素氧化酶I mt mRNA水平沿隐窝-绒毛轴相对均匀,但分级分离实验显示隐窝底部水平升高。与成熟绒毛肠上皮细胞相比,未成熟隐窝成肠细胞中的人类线粒体转录因子A水平也更高。这些结果表明,mt rRNA和mRNA的相对水平在肠道上皮细胞中根据细胞的隐窝-绒毛位置和分化状态受到明显调节,并且隐窝中的mt mRNA和mt TFA水平更高,这与未成熟成肠细胞线粒体生物发生过程中转录活性增加一致。