Resistance Gene-Guided Genome Mining: Serial Promoter Exchanges in Aspergillus nidulans Reveal the Biosynthetic Pathway for Fellutamide B, a Proteasome Inhibitor.
Resistance Gene-Guided Genome Mining: Serial Promoter Exchanges in Aspergillus nidulans Reveal the Biosynthetic Pathway for Fellutamide B, a Proteasome Inhibitor.
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DOI：
10.1021/acschembio.6b00213
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发表时间：
2016-08-19
影响因子：
4
通讯作者：
Oakley BR
中科院分区：
文献类型：
--
作者：
Yeh HH;Ahuja M;Chiang YM;Oakley CE;Moore S;Yoon O;Hajovsky H;Bok JW;Keller NP;Wang CC;Oakley BR
真菌基因组计划揭示了数千个隐藏的次级代谢（SM）生物合成基因簇，这些基因簇编码可能产生有价值化合物的途径。异源表达系统应该允许这些簇被表达并获得它们的产物，但是需要方法来鉴定最有价值的靶簇。构巢曲霉的inp簇包含编码蛋白酶体亚基的基因inpE，这使我们假设inp簇产生蛋白酶体抑制剂，并且inpE赋予对该化合物的抗性。先前表达该簇的努力失败了，但是通过用可调节的启动子顺序地替换簇的基因的启动子，我们成功地表达了它们。表达表明，该产品的inp集群是蛋白酶体抑制剂fellucinase B，我们的数据使我们能够提出一个生物合成途径的化合物。通过删除inpE和激活表达的inp集群，我们证明，inpE是需要内部产生的fellucidase B的抗性。这些数据提供了实验验证的假设，即一些真菌SM集群含有基因，编码的耐药形式的酶所针对的化合物产生的集群。
Fungal genome projects are revealing thousands of cryptic secondary metabolism (SM) biosynthetic gene clusters that encode pathways that potentially produce valuable compounds. Heterologous expression systems should allow these clusters to be expressed and their products obtained, but approaches are needed to identify the most valuable target clusters. The inp cluster of Aspergillus nidulans contains a gene, inpE, that encodes a proteasome subunit, leading us to hypothesize that the inp cluster produces a proteasome inhibitor and inpE confers resistance to this compound. Previous efforts to express this cluster have failed, but by sequentially replacing the promoters of the genes of the cluster with a regulatable promotor, we have expressed them successfully. Expression reveals that the product of the inp cluster is the proteasome inhibitor fellutamide B, and our data allow us to propose a biosynthetic pathway for the compound. By deleting inpE and activating expression of the inp cluster, we demonstrate that inpE is required for resistance to internally produced fellutamide B. These data provide experimental validation for the hypothesis that some fungal SM clusters contain genes that encode resistant forms of the enzymes targeted by the compound produced by the cluster.
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影响因子：
7.8
作者：
Chiang, Yi-Ming;Chang, Shu-Lin;Oakley, Berl R.;Wang, Clay C. C.
通讯作者：
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DOI：
10.1007/bf00339600
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期刊：
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作者：
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Yoshikawa, H
DOI：
10.1073/pnas.1205532110
发表时间：
2013-01-02
影响因子：
11.1
作者：
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通讯作者：
Mortensen, Uffe H.