Tang B, Nazar R N
Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.
J Biol Chem. 1992 Sep 5;267(25):17738-42.
A gene encoding the 5 S rRNA-binding protein (YL3) in yeast (Saccharomyces cerevisiae) was further characterized with respect to its chromosomal localization, the controlling sequence regions, and the influence of 5 S rRNA gene expression. Sequence and chromosome blot analyses localized the gene on chromosome XVI immediately downstream of a cytochrome oxidase assembly gene, COXII. S1 nuclease protection studies identified two major initiation sites, 20 and 65 nucleotides upstream of the coding sequence, and a single polyadenylation site, 98 nucleotides downstream of the stop codon. Northern blot analyses and S1 nuclease protection indicated a normal pattern of gene regulation in media supporting alternate rates of growth, but significantly unbalanced regulation was observed in the presence of mutant 5 S rRNA genes which under-produce RNA and result in reduced growth rates. The results suggest a co-ordinating regulatory mechanism which maintains appropriate levels of 5 S rRNA-protein complex; an internal control region-like sequence in the upstream region of the YL3 gene is consistent with this feedback mechanism.
对酵母(酿酒酵母)中编码5S核糖体RNA结合蛋白(YL3)的基因,就其染色体定位、控制序列区域以及对5S核糖体RNA基因表达的影响进行了进一步表征。序列分析和染色体印迹分析将该基因定位在第十六条染色体上,紧挨着细胞色素氧化酶组装基因COXII的下游。S1核酸酶保护研究确定了两个主要起始位点,位于编码序列上游20和65个核苷酸处,以及一个单一的聚腺苷酸化位点,位于终止密码子下游98个核苷酸处。Northern印迹分析和S1核酸酶保护表明,在支持不同生长速率的培养基中,基因调控模式正常,但在存在产生RNA不足并导致生长速率降低的突变型5S核糖体RNA基因的情况下,观察到明显的调控失衡。结果提示存在一种协调调控机制,可维持5S核糖体RNA-蛋白质复合物的适当水平;YL3基因上游区域类似内部控制区域的序列与这种反馈机制一致。