Shi Y G, Tyler B M
Department of Plant Pathology, University of California-Davis 95616.
Nucleic Acids Res. 1991 Dec 11;19(23):6511-7. doi: 10.1093/nar/19.23.6511.
The relative levels of rRNAs and ribosomal proteins are coordinately regulated by growth rate and carbon nutrition in Neurospora crassa. However, little is known about the mechanisms involved. To investigate the transcriptional regulation of ribosomal protein genes in N. crassa, we cloned and sequenced a ribosomal protein gene (crp-3). The inferred crp-3 protein sequence shares 89% and 83% homology at its N-terminus with the yeast rp51 and the human S17 ribosomal proteins respectively. The crp-3 gene contains two introns, neither of which are conserved in position with the RP51 or the S17 genes. The crp-3 gene is present in a single copy and was mapped by RFLP analysis to the right arm of linkage group IV, near the cot-1 locus. Sequence comparisons of the upstream regions of the three sequenced crp genes revealed several common features. These include a 'Taq box' (consensus: ARTTYGACTT) at -39, a CG repeat (consensus: CCCRCCRRR) at -65, and a major transcription initiation site embedded in a purine rich region flanked by an upstream pyrimidine rich sequence. Using four N.crassa ribosomal protein genes as probes, we demonstrated that the levels of the four ribosomal protein mRNAs were closely coordinated during a nutritional downshift from sucrose to quinic acid.
粗糙脉孢菌中rRNAs和核糖体蛋白的相对水平受生长速率和碳营养的协同调控。然而,其中涉及的机制却鲜为人知。为了研究粗糙脉孢菌中核糖体蛋白基因的转录调控,我们克隆并测序了一个核糖体蛋白基因(crp - 3)。推断出的crp - 3蛋白序列在其N端与酵母rp51和人类S17核糖体蛋白分别具有89%和83%的同源性。crp - 3基因包含两个内含子,其位置在RP51或S17基因中均不保守。crp - 3基因以单拷贝形式存在,并通过RFLP分析定位到连锁群IV的右臂,靠近cot - 1位点。对三个已测序的crp基因上游区域的序列比较揭示了几个共同特征。这些特征包括位于-39处的一个“Taq框”(共有序列:ARTTYGACTT)、位于-65处的一个CG重复序列(共有序列:CCCRCCRRR)以及一个位于富含嘌呤区域内的主要转录起始位点,该区域两侧为富含嘧啶的上游序列。使用四个粗糙脉孢菌核糖体蛋白基因作为探针,我们证明了在从蔗糖到奎尼酸的营养物质转换过程中,这四个核糖体蛋白mRNA的水平密切协调。