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Genome-wide analysis of transcriptional dependence and probable target sites for Abf1 and Rap1 in Saccharomyces cerevisiae

机译:全基因组分析的酿酒酵母中的转录依赖性和Abf1和Rap1可能的目标位点。

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Abf1 and Rap1 are general regulatory factors (GRFs) that contribute to transcriptional activation of a large number of genes, as well as to replication, silencing and telomere structure in yeast. In spite of their widespread roles in transcription, the scope of their functional targets genome-wide has not been previously determined. Here, we use microarrays to examine the contribution of these essential GRFs to transcription genome-wide, by using ts mutants that dissociate from their binding sites at 37 degrees C. We then combine this data with published ChIP-chip studies and motif analysis to identify probable direct targets for Abf1 and Rap1. We also identify a substantial number of genes likely to bind Rap1 or Abf1, but not affected by loss of GRF binding. Interestingly, the results strongly suggest that Rap1 can contribute to gene activation from farther upstream than can Abf1. Also, consistent with previous work, more genes that bind Abf1 are unaffected by loss of binding than those that bind Rap1. Finally, we show for several such genes that the Abf1 C-terminal region, which contains the putative activation domain, is not needed to confer this peculiar 'memory effect' that allows continued transcription after loss of Abf1 binding.
机译:Abf1和Rap1是一般调节因子(GRF),它们有助于大量基因的转录激活以及酵母中的复制,沉默和端粒结构。尽管它们在转录中具有广泛的作用,但以前尚未确定其功能靶标在全基因组范围内的范围。在这里,我们使用微阵列来检查这些必需的GRF对全基因组转录的贡献,方法是使用ts突变体,在37摄氏度时从其结合位点解离。然后将这些数据与已发表的ChIP芯片研究和基序分析相结合,以鉴定Abf1和Rap1的可能直接目标。我们还确定了可能与Rap1或Abf1结合但不受GRF结合丢失影响的大量基因。有趣的是,这些结果强烈表明,Rap1可以比Abf1促进上游基因激活。此外,与以前的工作一致,与结合Abf1的基因相比,结合Abf1的基因不受结合丢失的影响更大。最后,我们显示了几个这样的基因,不需要Abf1 C末端区域(包含推定的激活结构域)来赋予这种独特的“记忆效应”,从而在Abf1结合丧失后允许继续转录。

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