Genes for production of the enediyne antitumor antibiotic C-1027 in Streptomyces globisporus are clustered with the cagA gene that encodes the C-1027 apoprotein
Genes for production of the enediyne antitumor antibiotic C-1027 in Streptomyces globisporus are clustered with the cagA gene that encodes the C-1027 apoprotein
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球孢链霉菌中生产烯啶抗肿瘤抗生素 C-1027 的基因与编码 C-1027 蛋白的 cagA 基因聚集在一起
DOI：
10.1128/aac.44.2.382-392.2000
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发表时间：
2000-02-01
影响因子：
4.9
通讯作者：
Shen, B
中科院分区：
文献类型：
--
作者：
Liu, W;Shen, B
C-1027是烯二炔类抗肿瘤抗生素家族中最有效的成员，由球孢链霉菌C-1027产生，由一个脱辅蛋白(由cagA基因编码)和一个非肽类发色团组成。C-1027发色团可以被认为是由苯并恶唑酸酯、脱氧氨基己糖、对氨基酸和烯二炔核生物合成而来的。采用克隆C-1027生物合成基因簇的策略，将一个可能的dNDP-葡萄糖4，6-脱水酶基因定位到cagA上，定位了球孢链霉菌75kb的连续DNA。对克隆的基因簇的两个区域的DNA序列分析显示，两个基因sgcA和sgcB分别编码NGDH酶和跨膜外排蛋白，并证实cagA基因位于sgcAB基因座上游约14kb。通过破坏sgcA基因产生不产生C-1027的突变体，以及通过在体内补充sgcA突变体以恢复C-1027的生产，证明了克隆的基因簇参与了C-1027的生物合成，这些结果首次克隆了烯二炔抗肿瘤抗生素生物合成的基因簇，为未来C-1027生物合成的遗传和生化研究提供了一个起点。
C-1027, the most potent member of the enediyne antitumor antibiotic family, is produced by Streptomyces globisporus C-1027 and consists of an apoprotein (encoded by the cagA gene) and a nonpeptidic chromophore. The C-1027 chromophore could be viewed as being derived biosynthetically from a benzoxazolinate, a deoxy-amino hexose, a p-amino acid, and an enediyne core. By adopting a strategy for cloning of the C-1027 biosynthesis gene cluster by mapping a putative dNDP-glucose 4,6-dehydratase (NGDH) gene to cagA, we have localized 75 kb of contiguous DNA from S. globisporus. DNA sequence analysis of two regions of the cloned gene cluster revealed two genes, sgcA and sgcB, that encode an NGDH enzyme and a transmembrane efflux protein, respectively, and confirmed that the cagA gene resides approximately 14 kb upstream of the sgcAB locus. The involvement of the cloned gene cluster in C-1027 biosynthesis was demonstrated by disrupting the sgcA gene to generate C-1027-nonproducing mutants and by complementing the sgcA mutants in vivo to restore C-1027 production, These results represent the first cloning of a gene cluster for enediyne antitumor antibiotic biosynthesis and provide a starting point for future genetic and biochemical investigations of C-1027 biosynthesis.