Resistance Gene-Guided Genome Mining: Serial Promoter Exchanges in Aspergillus nidulans Reveal the Biosynthetic Pathway for Fellutamide B, a Proteasome Inhibitor

被引：87

作者：

Yeh, Hsu-Hua ^{[1
,7
]}

Ahuja, Manmeet ^{[2
,8
]}

Chiang, Yi-Ming ^{[1
,3
]}

Oakley, C. Elizabeth ^{[2
]}

Moore, Shauna ^{[2
]}

Yoon, Olivia ^{[2
]}

Hajoysky, Heather ^{[2
]}

Bok, Jin-Woo ^{[4
,5
]}

Keller, Nancy P. ^{[4
,5
]}

Wang, Clay C. C. ^{[1
,6
]}

Oakley, Berl R. ^{[2
]}

机构：

[1] Univ Southern Calif, Sch Pharm, Dept Pharmacol & Pharmaceut Sci, Los Angeles, CA 90089 USA

[2] Univ Kansas, Dept Mol Biosci, Lawrence, KS 66045 USA

[3] Chia Nan Univ Pharm & Sci, Dept Pharm, Tainan 71710, Taiwan

[4] Univ Wisconsin, Dept Bacteriol, Madison, WI 53706 USA

[5] Univ Wisconsin, Dept Med Microbiol & Immunol, Madison, WI 53706 USA

[6] Univ Southern Calif, Dept Chem, Dornsife Coll Letters Arts & Sci, Los Angeles, CA 90089 USA

[7] Chia Nan Univ Pharm & Sci, Drug Discovery & Dev Ctr, Tainan 71710, Taiwan

[8] Reliance Ind Ltd, Ind Biotechnol Div, Reliance Technol Grp, Reliance Corp Pk,Thane Belapur Rd, Ghansoli 400701, Navi Mumbia, India

来源：

ACS CHEMICAL BIOLOGY | 2016年 / 11卷 / 08期

关键词：

POLYKETIDE; CLUSTERS; FUNGUS; METABOLITES; SEQUENCE; SYSTEM;

D O I：

10.1021/acschembio.6b00213

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：2275 / 2284

页数：10