The posttranslational modification cascade to the thiopeptide berninamycin generates linear forms and altered macrocyclic scaffolds
The posttranslational modification cascade to the thiopeptide berninamycin generates linear forms and altered macrocyclic scaffolds
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DOI：
10.1073/pnas.1307111110
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发表时间：
2013-05-21
影响因子：
11.1
通讯作者：
Walsh, Christopher T.
中科院分区：
文献类型：
--
作者：
Malcolmson, Steven J.;Young, Travis S.;Walsh, Christopher T.
伯纳霉素是含吡啶硫肽类抗生素的一员，核糖体产生的前蛋白经过大量的翻译后修饰。Berninamycin的2-恶唑-3-噻唑-吡啶核心嵌入在35个原子的肽环中，而不是典型的三噻唑基吡啶核心嵌入在26个原子和29个原子的肽环中。作者从伯氏链霉菌UC 5144中克隆了一个含有11个基因的伯氏链霉菌簇，在lividans链霉菌TK24和委内瑞拉链霉菌ATCC 10712中进行了异源表达，并检测了变异和未完全加工的支架。野生型berninamycin前肽（BerA）和T3A突变体在S. lividans中翻译后成熟产生大环化合物和线性变异体，这些变异体无法形成吡啶和大环。该基因簇在委内瑞拉葡萄球菌中的表达产生了伯尼霉素35原子骨架的变体，在大环框架中含有甲基氯恶唑啉代替甲基氯恶唑。
Berninamycin is a member of the pyridine-containing thiopeptide class of antibiotics that undergoes massive posttranslational modifications from ribosomally generated preproteins. Berninamycin has a 2-oxazolyl-3-thiazolyl-pyridine core embedded in a 35-atom macrocycle rather than typical trithiazolylpyridine cores embedded in 26-atom and 29-atom peptide macrocycles. We describe the cloning of an 11-gene berninamycin cluster from Streptomyces bernensis UC 5144, its heterologous expression in Streptomyces lividans TK24 and Streptomyces venezuelae ATCC 10712, and detection of variant and incompletely processed scaffolds. Posttranslational maturation in S. lividans of both the wild-type berninamycin prepeptide (BerA) and also a T3A mutant generates macrocyclic compounds as well as linear variants, which have failed to form the pyridine and the macrocycle. Expression of the gene cluster in S. venezuelae generates a variant of the 35-atom skeleton of berninamycin, containing a methyloxazoline in the place of a methyloxazole within the macrocyclic framework.