Minvielle-Sebastia L, Winsor B, Bonneaud N, Lacroute F
Centre National de la Recherche Scientifique, Laboratoire propre associé à Université Pierre et Marie Curie, Gif-sur-Yvette, France.
Mol Cell Biol. 1991 Jun;11(6):3075-87. doi: 10.1128/mcb.11.6.3075-3087.1991.
In Saccharomyces cerevisiae, temperature-sensitive mutations in the genes RNA14 and RNA15 correlate with a reduction of mRNA stability and poly(A) tail length. Although mRNA transcription is not abolished in these mutants, the transcripts are rapidly deadenylated as in a strain carrying an RNA polymerase B(II) temperature-sensitive mutation. This suggests that the primary defect could be in the control of the poly(A) status of the mRNAs and that the fast decay rate may be due to the loss of this control. By complementation of their temperature-sensitive phenotype, we have cloned the wild-type genes. They are essential for cell viability and are unique in the haploid genome. The RNA14 gene, located on chromosome H, is transcribed as three mRNAs, one major and two minor, which are 2.2, 1.5, and 1.1 kb in length. The RNA15 gene gives rise to a single 1.2-kb transcript and maps to chromosome XVI. Sequence analysis indicates that RNA14 encodes a 636-amino-acid protein with a calculated molecular weight of 75,295. No homology was found between RNA14 and RNA15 or between RNA14 and other proteins contained in data banks. The RNA15 DNA sequence predicts a protein of 296 amino acids with a molecular weight of 32,770. Sequence comparison reveals an N-terminal putative RNA-binding domain in the RNA15-encoded protein, followed by a glutamine and asparagine stretch similar to the opa sequences. Both RNA14 and RNA15 wild-type genes, when cloned on a multicopy plasmid, are able to suppress the temperature-sensitive phenotype of strains bearing either the rna14 or the rna15 mutation, suggesting that the encoded proteins could interact with each other.
在酿酒酵母中,RNA14和RNA15基因的温度敏感型突变与mRNA稳定性降低及多聚腺苷酸(poly(A))尾长度缩短相关。尽管这些突变体中mRNA转录未被消除,但转录本会迅速去腺苷酸化,就如同携带RNA聚合酶B(II)温度敏感型突变的菌株一样。这表明主要缺陷可能在于对mRNA多聚腺苷酸状态的控制,而快速衰减率可能是由于这种控制的丧失。通过互补其温度敏感型表型,我们克隆了野生型基因。它们对细胞活力至关重要,且在单倍体基因组中是独特的。位于H染色体上的RNA14基因转录为三种mRNA,一种主要的和两种次要的,长度分别为2.2、1.5和1.1 kb。RNA15基因产生单一的1.2 kb转录本,定位于第十六号染色体。序列分析表明,RNA14编码一种636个氨基酸的蛋白质,计算分子量为75,295。在RNA14和RNA15之间或RNA14与数据库中包含的其他蛋白质之间未发现同源性。RNA15的DNA序列预测有一个296个氨基酸、分子量为32,770的蛋白质。序列比较显示,RNA15编码的蛋白质中有一个N端假定的RNA结合结构域,随后是一段类似于opa序列的谷氨酰胺和天冬酰胺延伸序列。当RNA14和RNA15野生型基因克隆到多拷贝质粒上时,它们都能够抑制携带rna14或rna15突变菌株的温度敏感型表型,这表明所编码的蛋白质可能相互作用。