Graphical analysis and experimental evaluation of Saccharomyces cerevisiae PTRK1|2 and PBMH1|2 promoter region.

Susanne Gerber, Guido Hasenbrink, Wouter Hendriksen, Paul Van Heusden, Jost Ludwig, Edda Klipp, Hella Lichtenberg-Fraté
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Abstract

We designed a simple graphical presentation for the results of a transcription factor (TF) pattern matching analysis. The TF analysis algorithm utilized known sequence signature motifs from several databases. The graphical presentation enabled a quick overview of potential TF binding sites, their frequency and spacing on both DNA strands and thus straight forward identification of promising candidates for further experimental investigations. The developed tool was applied on in total four Saccharomyces cerevisiae gene promoter regions. The selected differentially expressed genes belong to functionally different families and encode duplicate functions, TRK1 and TRK2 as ion transporters and BMH1 and BMH2 as multiple regulators. Output evaluation revealed a number of TFs with promising differences in the promoter regions of each gene pair. Experimental investigations were performed by using corresponding TF yeast mutants for either phenotypic analysis of ion transport mediated growth or expression analysis of BMH1,2 genes. Upon phenotypic testing one TF mutant exhibited severely impaired growth under non-permissive conditions. This TF, Mot3p was identified as of most abundant potential binding sites and distinctive patterns among the TRK promoter regions.

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酿酒酵母PTRK1|2和PBMH1|2启动子区图谱分析及实验评价
我们设计了一个简单的图形表示转录因子(TF)模式匹配分析的结果。TF分析算法利用了多个数据库中已知的序列特征基元。图形展示可以快速概述潜在的TF结合位点,它们在两条DNA链上的频率和间距,从而直接确定有希望的候选者进行进一步的实验研究。该工具应用于酿酒酵母基因启动子区域共4个。所选择的差异表达基因属于功能不同的家族,编码重复的功能,TRK1和TRK2作为离子转运体,BMH1和BMH2作为多重调节因子。输出评估显示,许多tf在每个基因对的启动子区域有希望的差异。实验研究采用相应的TF酵母突变体进行离子转运介导生长的表型分析或BMH1,2基因的表达分析。在表型测试中,一个TF突变体在非允许条件下表现出严重的生长受损。这个TF, Mot3p被认为是TRK启动子区域中最丰富的潜在结合位点和独特的模式。
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