Functional analysis of the yeast genome

Just as Saccharomyces cerevisiae itself provides a model for so many processes essential to eukaryotic life, we anticipate that the methods and the mindset that have moved yeast biological research "beyond the genome" provide a prototype for making similar progress in other organisms. In this review I describe the experimental processes, results and utility of the current large-scale experimental approaches that use genomic data to provide a functional analysis of the yeast genome. Electronic Publication     

Genome-wide identification,characterization and expression analysis of the LIM transcription factor family in quinoa

Lim family members play an important role in the regulation of plant cell development and stress response. However, there are few studies on LIM family in quinoa. In this study, we identified nine LIMS (named cqlim01-cqlim09) from quinoa, which were divided into three subfamilies (α Lim1, γ lim2 and δ lim2) according to phylogeny. The differences in gene structure and motif composition among different subfamilies have been observed. In addition, we studied the repetitive events of the members of the family. The Ka/Ks (non synchronous substitution rate / synchronous substitution rate) ratio analysis showed that the repetitive CqLIMs probably experienced purifying selection pressure. Promoter analysis showed that the family genes contained a variety of hormones, stress and tissue-specific expression elements, and protein interactions showed that these genes had actin stabilizing effect. In addition, QRT PCR results showed that all CqLIM genes were positively regulated under three stresses (low temperature, salt and ABA). These results provide a theoretical basis of further study of LIM gene in quinoa. Supplementary informationThe online version contains supplementary material available at 10.1007/s12298-021-00988-2.     

Two proteins act as the IUF1 insulin gene enhancer binding factor

IUF1 is a pancreatic β cell-specific factor which binds to the sequence 5′-CPyCTAATG-3′ (CT box) within the human insulin gene enhancer. Here we show that IUF1 is composed of 2 binding activities that can be separated by DEAE ion exchange chromatography. South Western blot analysis indicates that these distinct binding activities have apparent molecular weights of 115 kDa and 46 kDa.     

冠突散囊菌转录因子stf1基因的克隆及其表达分析

利用RACE技术获得了冠突散囊菌stf1基因的全长DNA序列。该序列全长3 029bp,开放阅读框长度为2 664bp,从114–2 908bp,在253bp处含有一个131bp的内含子,预测编码887个氨基酸。同源分析结果表明该基因与Snd1/p100转录因子同源。应用相对定量SYBR Green I荧光定量PCR技术对stf1基因在不同发育时期的表达量的差异进行了检测。结果表明,这个基因在子囊孢子时期的表达量最高,在分生孢子时期的表达量最低,相比子囊孢子时期下降了1倍。为深入研究冠突散囊菌的产孢调控机制奠定了一定的基础。     

Functional expression of the maize mitochondrial URF13 down-regulates galactose-induced GAL1 gene expression in Saccharomyces cerevisiae

Genes for the enzymes that metabolize galactose in Saccharomyces cerevisiae are strongly induced by galactose and tightly repressed by glucose. Because glucose also represses mitochondrial activity, we examined if derepression of the GAL1 galactokinase gene requires physiologically active mitochondria. The effect of mitochondria on the expression of GAL1 was analyzed by a novel approach in which the activity of the organelles was altered by functional expression of URF13, a mitochondrial protein unique to the Texas-type cytoplasmic male sterility phenotype in maize. Mitochondrial targeting and functional expression of the URF13 protein in yeast result in a decrease of the mitochondrial membrane potential similar to those observed in cells treated with mitochondrial inhibitors such as antimycin A or sodium azide. Activation of URF13 in galactose-induced cells results in the inhibition of GAL1 expression in the absence of repressing concentrations of glucose. Our data reveal the existence of a regulatory pathway that connects the derepression of the GAL1 gene with mitochondrial activity.