Dry and wet approaches for genome-wide functional annotation of conventional and unconventional transcriptional activators

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• 作者：Elisabetta Levati ; Sara Sartini ; Simone Ottonello ; ¹ ; ^{simone.ottonello@unipr.it} ; Barbara Montanini¹ ; ^{barbara.montanini@unipr.it}
• 关键词：Transcription factors ; Moonlighting transcriptional activators ; Transcriptional activator trap ; Yeast/bacterial one-hybrid ; Protein binding microarrays ; HT-SELEX
• 刊名：Computational and Structural Biotechnology Journal
• 出版年：2016
• 出版时间：2016
• 年：2016
• 卷：14
• 期：Complete
• 页码：262-270
• 全文大小：577 K
• 卷排序：14

文摘

Transcription factors (TFs) are master gene products that regulate gene expression in response to a variety of stimuli. They interact with DNA in a sequence-specific manner using a variety of DNA-binding domain (DBD) modules. This allows to properly position their second domain, called “effector domain”, to directly or indirectly recruit positively or negatively acting co-regulators including chromatin modifiers, thus modulating preinitiation complex formation as well as transcription elongation. At variance with the DBDs, which are comprised of well-defined and easily recognizable DNA binding motifs, effector domains are usually much less conserved and thus considerably more difficult to predict. Also not so easy to identify are the DNA-binding sites of TFs, especially on a genome-wide basis and in the case of overlapping binding regions. Another emerging issue, with many potential regulatory implications, is that of so-called “moonlighting” transcription factors, i.e., proteins with an annotated function unrelated to transcription and lacking any recognizable DBD or effector domain, that play a role in gene regulation as their second job. Starting from bioinformatic and experimental high-throughput tools for an unbiased, genome-wide identification and functional characterization of TFs (especially transcriptional activators), we describe both established (and usually well affordable) as well as newly developed platforms for DNA-binding site identification. Selected combinations of these search tools, some of which rely on next-generation sequencing approaches, allow delineating the entire repertoire of TFs and unconventional regulators encoded by the any sequenced genome.