Chen W, Dieckmann C L
Department of Biochemistry, University of Arizona, Tucson 85721.
J Biol Chem. 1994 Jun 17;269(24):16574-8.
Cbp1p is a nuclear encoded protein required for the stabilization of mitochondrial COB mRNA, which codes for apocytochrome b in the yeast Saccharomyces cerevisiae. The COB gene is cotranscribed with the upstream tRNA(Glu) gene. Release of tRNA(Glu) from the initial transcript generates a precursor mRNA with a 5'-end at position -1098. The 5'-end of mature COB message is generated by cleavage of the pre-mRNA at nucleotide -955 or -954. Previous work indicated that Cbp1p acts through cis-elements near these cleavage sites. Here we have tested whether Cbp1p stabilizes COB mRNA solely by stimulating the processing of COB precursor RNA at nucleotide -955/-954. Yeast strain TG955 was constructed such that the -955 COB mRNA 5'-processing site coincides with the upstream tRNA 3'-endonuclease site at position -1098, allowing the 5'-end of COB mRNA to be formed by the tRNA 3'-processing enzyme. Respiratory growth and stability of COB mRNA in TG955 are Cbp1p-dependent. Therefore, we conclude that Cbp1p is important for the stabilization of COB mRNA after 5'-processing.
Cbp1p是一种核编码蛋白,对于酿酒酵母中线粒体COB mRNA的稳定是必需的,COB mRNA编码脱辅基细胞色素b。COB基因与上游的tRNA(Glu)基因共转录。从初始转录本中释放tRNA(Glu)会产生一个5'端位于-1098位置的前体mRNA。成熟COB mRNA的5'端是通过在前体mRNA的-955或-954核苷酸处切割产生的。先前的工作表明,Cbp1p通过这些切割位点附近的顺式元件起作用。在这里,我们测试了Cbp1p是否仅通过刺激COB前体RNA在-955 / -954核苷酸处的加工来稳定COB mRNA。构建了酵母菌株TG955,使得-955 COB mRNA的5'加工位点与-1098位置的上游tRNA 3'内切核酸酶位点重合,从而使COB mRNA的5'端由tRNA 3'加工酶形成。TG955中COB mRNA的呼吸生长和稳定性依赖于Cbp1p。因此,我们得出结论,Cbp1p对于5'加工后COB mRNA的稳定很重要。