Gómez-Navarro Natalia, Jordán-Pla Antonio, Estruch Francisco, E Pérez-Ortín José
Departamento de Bioquímica y Biología Molecular, Facultad de Biológicas and ERI Biotecmed, Universitat de València, Dr. Moliner 50, E-46100, Burjassot, Valencia, Spain.
Present address: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
BMC Genomics. 2016 Mar 3;17:183. doi: 10.1186/s12864-016-2536-2.
The formation of the pre-initiation complex in eukaryotic genes is a key step in transcription initiation. The TATA-binding protein (TBP) is a universal component of all pre-initiation complexes for all kinds of RNA polymerase II (RNA pol II) genes, including those with a TATA or a TATA-like element, both those that encode proteins and those that transcribe non-coding RNAs. Mot1 and the negative cofactor 2 (NC2) complex are regulators of TBP, and it has been shown that depletion of these factors in yeast leads to defects in the control of transcription initiation that alter cryptic transcription levels in selected yeast loci.
In order to cast light on the molecular functions of NC2, we performed genome-wide studies in conditional mutants in yeast NC2 essential subunits Ydr1 and Bur6. Our analyses show a generally increased level of cryptic transcription in all kinds of genes upon depletion of NC2 subunits, and that each kind of gene (canonical or ncRNAs, TATA or TATA-like) shows some differences in the cryptic transcription pattern for each NC2 mutant.
We conclude that NC2 plays a general role in transcription initiation in RNA polymerase II genes that is related with its known TBP interchange function from free to promoter bound states. Therefore, loss of the NC2 function provokes increases in cryptic transcription throughout the yeast genome. Our results also suggest functional differences between NC2 subunits Ydr1 and Bur6.
真核基因中起始前复合物的形成是转录起始的关键步骤。TATA结合蛋白(TBP)是所有RNA聚合酶II(RNA pol II)基因的所有起始前复合物的通用组成部分,包括那些具有TATA或TATA样元件的基因,无论是编码蛋白质的基因还是转录非编码RNA的基因。Mot1和负辅因子2(NC2)复合物是TBP的调节因子,并且已经表明,在酵母中耗尽这些因子会导致转录起始控制缺陷,从而改变选定酵母基因座中的隐蔽转录水平。
为了阐明NC2的分子功能,我们对酵母NC2必需亚基Ydr1和Bur6的条件突变体进行了全基因组研究。我们的分析表明,在耗尽NC2亚基后,各类基因中的隐蔽转录水平普遍增加,并且每种基因(经典基因或非编码RNA基因、TATA基因或TATA样基因)在每个NC2突变体的隐蔽转录模式上都存在一些差异。
我们得出结论,NC2在RNA聚合酶II基因的转录起始中起普遍作用,这与其已知的从游离状态到启动子结合状态的TBP交换功能有关。因此,NC2功能的丧失会导致整个酵母基因组中隐蔽转录的增加。我们的结果还表明NC2亚基Ydr1和Bur6之间存在功能差异。