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真菌卷枝毛霉中的线粒体柠檬酸转运系统:鉴定、系统发育分析和生长及脂质积累过程中的表达谱分析。

Mitochondrial Citrate Transport System in the Fungus Mucor circinelloides: Identification, Phylogenetic Analysis, and Expression Profiling During Growth and Lipid Accumulation.

机构信息

Colin Ratledge Center for Microbial Lipids, School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo, 255000, Shandong, People's Republic of China.

Institute of Biotechnology, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinskeho 9, 812 37, Bratislava, Slovakia.

出版信息

Curr Microbiol. 2020 Feb;77(2):220-231. doi: 10.1007/s00284-019-01822-5. Epub 2019 Dec 4.

Abstract

The mitochondrial citrate transport system, composed of citrate and malate transporters (MTs), can regulate the citrate efflux from mitochondria to cytosol, and then citrate is cleaved into OAA and acetyl-CoA which can be used for fatty acid (FA) biosynthesis. However, in the fungus Mucor circinelloides the molecular mechanism of citrate efflux from the mitochondria by this system and its role in FA synthesis is unclear. In the present study, we have analyzed the genome of high lipid-producing strain WJ11 and the low lipid-producing strain CBS 277.49 to find the potential genes involving in this system. Five potential genes are present in the genome of WJ11. These genes encode one citrate transport protein (CT), one tricarboxylate carrier (TCT), one MT, and two 2-oxoglutarate:malate antiporters (SoDIT-a and SoDIT-b). However, the genome of CBS 277.49 contains the same set of genes, except for the presence of just one SoDIT. The proteins from WJ11 had similar properties as their counterparts in CBS 277.49. Moreover, phylogenetic analyses revealed the evolutionary relationship of these proteins and illuminated their typical motifs related to potential functions. Additionally, the expression of these genes was analyzed to predict the possible functions in lipid metabolism in M. circinelloides. This is the first study to report the in silico analysis of structures and functions of the mitochondrial citrate transport system in M. circinelloides. This work showed a new strategy for research for the selection of candidate genes for further detailed functional investigation of the mitochondrial citrate transport system in lipid accumulation.

摘要

线粒体柠檬酸转运系统由柠檬酸和苹果酸转运蛋白(MTs)组成,可调节柠檬酸从线粒体向细胞质的外排,然后柠檬酸被裂解为草酰乙酸(OAA)和乙酰辅酶 A,可用于脂肪酸(FA)合成。然而,在真菌卷枝毛霉中,该系统的柠檬酸从线粒体外排的分子机制及其在 FA 合成中的作用尚不清楚。在本研究中,我们分析了高产脂菌株 WJ11 和低产脂菌株 CBS 277.49 的基因组,以寻找可能涉及该系统的潜在基因。WJ11 基因组中存在 5 个潜在基因。这些基因编码一个柠檬酸转运蛋白(CT)、一个三羧酸载体(TCT)、一个 MT 和两个 2-氧戊二酸:苹果酸反向转运蛋白(SoDIT-a 和 SoDIT-b)。然而,CBS 277.49 的基因组包含相同的一组基因,只是缺少一个 SoDIT。WJ11 的蛋白具有与 CBS 277.49 中的蛋白相似的特性。此外,系统发育分析揭示了这些蛋白的进化关系,并阐明了它们与潜在功能相关的典型基序。此外,还分析了这些基因的表达,以预测它们在卷枝毛霉脂质代谢中的可能功能。这是首次报道卷枝毛霉线粒体柠檬酸转运系统结构和功能的计算机分析。这项工作为进一步详细研究线粒体柠檬酸转运系统在脂质积累中的功能提供了一种新的候选基因选择策略。

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