Morales M J, Dang Y L, Lou Y C, Sulo P, Martin N C
Department of Biochemistry, University of Louisville, KY 40292.
Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9875-9. doi: 10.1073/pnas.89.20.9875.
RNase P from the mitochondria of Saccharomyces cerevisiae was purified to near homogeneity > 1800-fold with a yield of 1.6% from mitochondrial extracts. The most abundant protein in the purified fractions is, at 105 kDa, considerably larger than the 14-kDa bacterial RNase P protein subunits. Oligonucleotides designed from the amino-terminal sequence of the 105-kDa protein were used to identify and isolate the 105-kDa protein-encoding gene. Strains carrying a disruption of the gene for the 105-kDa protein are viable but respiratory deficient and accumulate mitochondrial tRNA precursors with 5' extensions. As this is the second gene known to be necessary for yeast mitochondrial RNase P activity, we have named it RPM2 (for RNase P mitochondrial).
从酿酒酵母线粒体中纯化出的核糖核酸酶P(RNase P)达到了近乎均一的状态,比初始线粒体提取物纯化了1800倍以上,产率为1.6%。纯化组分中最丰富的蛋白质分子量为105 kDa,比14 kDa的细菌RNase P蛋白质亚基大得多。根据105 kDa蛋白质的氨基末端序列设计的寡核苷酸被用于鉴定和分离编码该105 kDa蛋白质的基因。携带105 kDa蛋白质基因缺失的菌株能够存活,但呼吸功能有缺陷,并且会积累带有5'端延伸的线粒体tRNA前体。由于这是已知对酵母线粒体RNase P活性必需的第二个基因,我们将其命名为RPM2(线粒体核糖核酸酶P相关蛋白2)。