Lee F J, Moss J
Laboratory of Cellular Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
J Biol Chem. 1993 Jul 15;268(20):15080-7.
A gene, termed RNA-binding protein (RBP1), was cloned from Saccharomyces cerevisiae. RBP1 contains an open reading frame of 2016 nucleotides that encodes a 672-amino acid protein with a calculated M(r) of approximately 75,000. Southern blots of genomic DNA from wild-type and RBP1-disrupted strains were consistent with the presence of homologous genes. RNA blots revealed a major 2.7-kb RNA band and two minor bands of 1.5 and 1.1 kb. The sequence of the putative RBP1 protein contains two copies of an RNA recognition motif, two glutamine stretches, an asparagine-rich region, a methionine-rich region, and two long potential alpha-helixes. In addition, recombinant RBP1 fusion protein can bind to RNA and single-stranded DNA but not double-stranded DNA. RBP1 is a glucose-repressible gene. Disruption of RBP1 increased cell growth rate in the early log phase. Overexpression of RBP1 or reduction in its translation by expression of antisense RNA decreased or increased the cell growth rate, respectively. From these observations, we infer that RBP1 may be involved in growth regulation, possibly through its participation in RNA metabolism.
从酿酒酵母中克隆出一个名为RNA结合蛋白(RBP1)的基因。RBP1含有一个2016个核苷酸的开放阅读框,编码一个672个氨基酸的蛋白质,计算所得的分子量约为75,000。野生型和RBP1缺失菌株基因组DNA的Southern杂交结果与同源基因的存在相符。RNA印迹显示有一条主要的2.7kb RNA条带以及两条1.5kb和1.1kb的较小条带。推测的RBP1蛋白序列包含两个RNA识别基序拷贝、两个谷氨酰胺延伸段、一个富含天冬酰胺的区域、一个富含甲硫氨酸的区域以及两个长的潜在α螺旋。此外,重组RBP1融合蛋白可与RNA和单链DNA结合,但不与双链DNA结合。RBP1是一个可被葡萄糖抑制的基因。RBP1的缺失在对数生长期早期提高了细胞生长速率。RBP1的过表达或通过反义RNA表达降低其翻译水平,分别降低或提高了细胞生长速率。基于这些观察结果,我们推断RBP1可能参与生长调节,可能是通过参与RNA代谢来实现的。