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Dal82p结构域在酿酒酵母中脲基甲酸酯/草尿酸盐依赖性基因表达中的作用。

Roles of the Dal82p domains in allophanate/oxalurate-dependent gene expression in Saccharomyces cerevisiae.

作者信息

Scott S, Abul-Hamd A T, Cooper T G

机构信息

Department of Microbiology and Immunology, University of Tennessee, Memphis, Tennessee 38163, USA.

出版信息

J Biol Chem. 2000 Oct 6;275(40):30886-93. doi: 10.1074/jbc.M005624200.

DOI:10.1074/jbc.M005624200
PMID:10906145
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4382018/
Abstract

Allophanate/oxalurate-induced gene expression in Saccharomyces cerevisiae requires at least five transcription factors, four of which act positively (Gln3p, Gat1p, Dal81p, and Dal82p) and one negatively (Dal80p). Gln3p binds to and Gat1p is proposed to bind to single GATA sequences; Dal80p binds to pairs of specifically oriented and spaced GATA sequences, and Dal82p binds to a pathway-specific element, UIS(ALL). Dal82p consists of at least three domains as follows: (i) UIS(ALL) DNA-binding, (ii) transcriptional activation, and (iii) coiled-coil(DAL82). Here we show that the coiled-coil(DAL82) domain possesses two demonstrable functions. (i) It prevents Dal82p-mediated transcription when inducer is absent. (ii) It is a major, although not exclusive, domain through which the inducer signal is received. Supporting the latter conclusion, a 38-amino acid fragment, containing little more than the coiled-coil(DAL82) domain, supports oxalurate-inducible, Dal81p-dependent, reporter gene transcription. Dal81p is required for inducer responsiveness of LexAp-Dal82p and LexAp coiled-coil(DAL82)-mediated transcription but isn't needed for inducer-dependent activation mediated by a Dal82p containing deletions in both the coiled-coil(DAL82), UIS(ALL)-binding domains. There may be an interaction between Dal81p and the coiled-coil(DAL82) domain since (i) Dal81p is required for transcription mediated by LexA-coiled-coil(DAL82)p and (ii) a Dal81p-Dal82p complex is detected by two-hybrid assay.

摘要

脲基甲酸酯/草尿酸酯诱导的酿酒酵母基因表达至少需要五种转录因子，其中四种起正向作用（Gln3p、Gat1p、Dal81p和Dal82p），一种起负向作用（Dal80p）。Gln3p可结合，并且推测Gat1p可结合单个GATA序列；Dal80p结合特定方向和间隔的GATA序列对，而Dal82p结合一条途径特异性元件UIS(ALL)。Dal82p至少由三个结构域组成：(i) UIS(ALL) DNA结合结构域，(ii) 转录激活结构域，以及(iii) 卷曲螺旋（DAL82）结构域。在此我们表明卷曲螺旋（DAL82）结构域具有两种可证实的功能。(i) 在没有诱导剂时，它可阻止Dal82p介导的转录。(ii) 它是接收诱导剂信号的主要结构域，尽管不是唯一的结构域。支持后一结论的是，一个仅比卷曲螺旋（DAL82）结构域多一点的38个氨基酸的片段可支持草尿酸酯诱导的、Dal81p依赖的报告基因转录。LexAp-Dal82p和LexAp卷曲螺旋（DAL82）介导的转录的诱导剂反应性需要Dal81p，但由在卷曲螺旋（DAL82）和UIS(ALL)结合结构域均有缺失的Dal82p介导的诱导剂依赖性激活则不需要Dal81p。由于(i) LexA-卷曲螺旋（DAL82）p介导的转录需要Dal81p，以及(ii) 通过双杂交分析检测到Dal81p-Dal82p复合物，因此Dal81p与卷曲螺旋（DAL82）结构域之间可能存在相互作用。