Mahadevan S, Struhl K
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
Mol Cell Biol. 1990 Sep;10(9):4447-55. doi: 10.1128/mcb.10.9.4447-4455.1990.
Tc is the proximal promoter element required for constitutive his3 transcription that occurs in the absence of the canonical TATA element (TR) and is initiated from the +1 site. The TC element, unlike TR, does not respond to transcriptional stimulation by the GCN4 or GAL4 activator protein. Analysis of deletion, substitution, and point mutations indicates that Tc mapped between nucleotides -54 and -83 and is a sequence-dependent element because it could not be functionally replaced by other DNA sequences. However, in contrast to the behavior of typical promoter elements, it was surprisingly difficult to eliminate Tc function by base pair substitutions. Of 15 derivatives averaging four substitutions in the Tc region and representing 40% of all possible single changes, only 1 inactivated the Tc element. Moreover, the phenotypes of mutant and hybrid elements indicated that inactivation of Tc required multiple changes. The spacing between Tc and the initiation region could be varied over a 30-base-pair range without significantly affecting the level of transcription from the +1 site. From these results, we consider it possible that Tc may not interact with TFIID or some other typical sequence-specific transcription factor, but instead might influence transcription, either directly or indirectly, by its DNA structure.
Tc是组成型his3转录所需的近端启动子元件,该转录在没有典型TATA元件(TR)的情况下发生,并从+1位点起始。与TR不同,TC元件不响应GCN4或GAL4激活蛋白的转录刺激。缺失、替换和点突变分析表明,Tc定位于核苷酸-54至-83之间,是一个序列依赖性元件,因为它不能被其他DNA序列功能性取代。然而,与典型启动子元件的行为相反,通过碱基对替换消除Tc功能出奇地困难。在Tc区域平均有四个替换且代表所有可能单碱基变化40%的15个衍生物中,只有1个使Tc元件失活。此外,突变和杂交元件的表型表明,Tc失活需要多个变化。Tc与起始区域之间的间距可以在30个碱基对的范围内变化,而不会显著影响从+1位点的转录水平。根据这些结果,我们认为Tc可能不与TFIID或其他一些典型的序列特异性转录因子相互作用,而是可能通过其DNA结构直接或间接地影响转录。