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来自酿酒酵母的生物素合酶基因:大肠杆菌生物素合酶缺失菌株的克隆、测序及互补作用

The gene for biotin synthase from Saccharomyces cerevisiae: cloning, sequencing, and complementation of Escherichia coli strains lacking biotin synthase.

作者信息

Zhang S, Sanyal I, Bulboaca G H, Rich A, Flint D H

机构信息

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

出版信息

Arch Biochem Biophys. 1994 Feb 15;309(1):29-35. doi: 10.1006/abbi.1994.1079.

Abstract

Biotin synthase catalyzes the insertion of a sulfur atom between two carbon atoms of dethiobiotin to form biotin in the last step of the biotin biosynthesis pathway. In Escherichia coli, biotin synthase is coded for by bioB gene. We report here cloning, sequencing, and initial functional characterization of the yeast gene for biotin synthase in Saccharomyces cerevisiae. We have named this gene BIO2. It consists of a 355-codon open reading frame near the ZUO1 gene. Analysis of the yeast protein encoded by the BIO2 gene reveals that it shares extensive homology with biotin synthases of E. coli and Bacillus sphaericus. The yeast and the two bacterial biotin synthase proteins have similar molecular weights, amino acid compositions, and hydropathies. The plasmid pUCBIO2 containing the yeast BIO2 gene completely complements E. coli bioB- and delta bio mutants and enables these mutants to grow on dethiobiotin. Although BIO2 is physically linked to ZUO1, which encodes the putative left-handed Z-DNA binding protein zuotin, it appears to be regulated independently from it. The yeast BIO2 and ZUO1 genes reside near ADE3 gene on chromosome VII. BIO2 is the first eukaryotic gene reported from the biotin biosynthetic pathway.

摘要

生物素合酶在生物素生物合成途径的最后一步催化硫原子插入脱硫生物素的两个碳原子之间以形成生物素。在大肠杆菌中,生物素合酶由bioB基因编码。我们在此报告酿酒酵母中生物素合酶酵母基因的克隆、测序及初步功能表征。我们将此基因命名为BIO2。它由位于ZUO1基因附近的一个355个密码子的开放阅读框组成。对由BIO2基因编码的酵母蛋白的分析表明,它与大肠杆菌和球形芽孢杆菌的生物素合酶具有广泛的同源性。酵母以及这两种细菌的生物素合酶蛋白具有相似的分子量、氨基酸组成和亲水性。含有酵母BIO2基因的质粒pUCBIO2完全互补大肠杆菌bioB-和δbio突变体,并使这些突变体能够在脱硫生物素上生长。尽管BIO2与编码假定的左手Z-DNA结合蛋白zuotin的ZUO1在物理上相连,但它似乎是独立于ZUO1进行调控的。酵母BIO2和ZUO1基因位于第七条染色体上的ADE3基因附近。BIO2是生物素生物合成途径中报道的第一个真核基因。

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