Henry Y A, López M C, Gibbs J M, Chambers A, Kingsman S M, Baker H V, Stanway C A
Department of Biochemistry, Oxford University, UK.
Mol Gen Genet. 1994 Nov 15;245(4):506-11. doi: 10.1007/BF00302263.
Analysis of the upstream activation sequence (UAS) of the yeast phosphoglycerate kinase gene (PGK) has demonstrated that a number of sequence elements are involved in its activity and two of these sequences are bound by the multifunctional factors Rap1p and Abf1p. In this report we show by in vivo footprinting that the regulatory factor encoded by GCR1 binds to two elements in the 3' half of the PGK UAS. These elements contain the sequence CTTCC, which was previously suggested to be important for the activity of the PGK UAS and has been shown to be able to bind Gcr1p in vitro. Furthermore, we find that Gcr1p positively influences PGK transcription, although it is not responsible for the carbon source dependent regulation of PGK mRNA synthesis. In order to mediate its transcriptional influence we find that Gcr1p requires the Rap1p binding site, in addition to its own, but not the Abf1p site. As neither a Rap1p nor a Gcr1p binding site alone is able to activate transcription, we propose that Gcr1p and Rap1p interact in an interdependent fashion to activate PGK transcription.
对酵母磷酸甘油酸激酶基因(PGK)上游激活序列(UAS)的分析表明,许多序列元件参与其活性,其中两个序列与多功能因子Rap1p和Abf1p结合。在本报告中,我们通过体内足迹法表明,由GCR1编码的调节因子与PGK UAS 3' 端的两个元件结合。这些元件包含序列CTTCC,先前认为该序列对PGK UAS的活性很重要,并且已证明在体外能够结合Gcr1p。此外,我们发现Gcr1p对PGK转录有正向影响,尽管它不负责PGK mRNA合成的碳源依赖性调节。为了介导其转录影响,我们发现Gcr1p除了自身的结合位点外,还需要Rap1p结合位点,但不需要Abf1p位点。由于单独的Rap1p结合位点或Gcr1p结合位点都不能激活转录,我们提出Gcr1p和Rap1p以相互依赖的方式相互作用以激活PGK转录。