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编码一种与植物角质诱导真菌角质酶所必需的回文启动子元件结合的蛋白质的cDNA的克隆与表达

Cloning and expression of cDNA encoding a protein that binds a palindromic promoter element essential for induction of fungal cutinase by plant cutin.

作者信息

Li D, Kolattukudy P E

机构信息

Department of Neurobiotechnology, Ohio State University, Columbus 43210, USA.

出版信息

J Biol Chem. 1995 May 19;270(20):11753-6. doi: 10.1074/jbc.270.20.11753.

Abstract

Previous studies showed that a palindromic sequence located at -159 base pairs is essential for induction of cutinase gene in Fusarium solani f. sp. pisi (Nectria haematococca mating type VI) by the hydroxy fatty acids from plant cutin and that a 50-kDa nuclear protein binds to a promoter that contains this element. Screening of a phage lambda gt11 expression library with the concatenated palindromic sequence as the probe identified a cDNA encoding a palindrome-binding protein (PBP). Nucleotide sequence of this cDNA revealed an open reading frame that would code for PBP with a calculated molecular weight of 49,847. This PBP contains a putative nuclear localization signal and a zinc finger motif sharing homology with the zinc finger DNA binding domains of transcription factors from mammals, Saccharomyces cerevisiae, Neurospora crassa, and Ustilago maydis. A highly basic region immediately adjacent to the carboxyl side of the zinc finger was also observed. PBP expressed in Escherichia coli showed specific binding to the palindromic DNA fragment.

摘要

先前的研究表明,位于-159个碱基对处的回文序列对于茄丝核菌(豌豆专化型,血红色球座菌交配型VI)中角质酶基因被来自植物角质的羟基脂肪酸诱导表达至关重要,并且一种50 kDa的核蛋白可与含有该元件的启动子结合。用串联的回文序列作为探针筛选λ噬菌体gt11表达文库,鉴定出一个编码回文结合蛋白(PBP)的cDNA。该cDNA的核苷酸序列揭示了一个开放阅读框,其编码的PBP计算分子量为49,847。这种PBP含有一个假定的核定位信号和一个锌指基序,与来自哺乳动物、酿酒酵母、粗糙脉孢菌和玉米黑粉菌的转录因子的锌指DNA结合结构域具有同源性。在锌指羧基侧紧邻处还观察到一个高度碱性区域。在大肠杆菌中表达的PBP显示出与回文DNA片段的特异性结合。

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