酿酒酵母中α-异丙基苹果酸合成酶的反馈调控和异寡聚调控。

Eukaryotic Cell Pub Date : 2015-06-01 Epub Date: 2015-04-03 DOI:10.1128/EC.00033-15
Geovani López, Héctor Quezada, Mariana Duhne, James González, Mijail Lezama, Mohammed El-Hafidi, Maritrini Colón, Ximena Martínez de la Escalera, Mirelle Citlali Flores-Villegas, Claudio Scazzocchio, Alexander DeLuna, Alicia González
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引用次数: 19

摘要

α-异丙基苹果酸酯(α-IPM)的产生对亮氨酸的生物合成和代谢的全局控制至关重要。出芽酵母(Saccharomyces cerevisiae)具有编码α-IPM合成酶(α-IPMS)同工酶的两个同源基因LEU4和LEU9。这两种α-IPMS同工酶之间的生化差异尚不清楚。在这里,我们发现Leu4同二聚体是亮氨酸敏感的异构体,而Leu9同二聚体对这种反馈抑制具有抗性。leu4Δ突变体只表达抗反馈的亮氨酸9同二聚体,在葡萄糖或乙醇的作用下生长缓慢,积累了大量亮氨酸;这种表型通过添加亮氨酸而得到缓解。用携带LEU4的着丝粒质粒转化leu4Δ突变体恢复了野生型表型。双分子荧光互补分析表明,在体内形成了Leu4-Leu9异二聚体同工酶。纯化和动力学分析表明,该异质寡聚同工酶具有明显的亮氨酸敏感性。在乙醇培养物中α-IPMS活性的测定表明,在这种呼吸条件下α-IPM的生物合成和生长依赖于反馈敏感的Leu4同二聚体。我们得出结论,两种酵母α- ipms的保留和进一步多样化导致了一个特定的调节系统,该系统通过对同质和异二聚体异构体的选择性反馈敏感性来控制亮氨酸-α-IPM生物合成途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Diversification of Paralogous α-Isopropylmalate Synthases by Modulation of Feedback Control and Hetero-Oligomerization in Saccharomyces cerevisiae.

Diversification of Paralogous α-Isopropylmalate Synthases by Modulation of Feedback Control and Hetero-Oligomerization in Saccharomyces cerevisiae.

Diversification of Paralogous α-Isopropylmalate Synthases by Modulation of Feedback Control and Hetero-Oligomerization in Saccharomyces cerevisiae.

Diversification of Paralogous α-Isopropylmalate Synthases by Modulation of Feedback Control and Hetero-Oligomerization in Saccharomyces cerevisiae.

Production of α-isopropylmalate (α-IPM) is critical for leucine biosynthesis and for the global control of metabolism. The budding yeast Saccharomyces cerevisiae has two paralogous genes, LEU4 and LEU9, that encode α-IPM synthase (α-IPMS) isozymes. Little is known about the biochemical differences between these two α-IPMS isoenzymes. Here, we show that the Leu4 homodimer is a leucine-sensitive isoform, while the Leu9 homodimer is resistant to such feedback inhibition. The leu4Δ mutant, which expresses only the feedback-resistant Leu9 homodimer, grows slowly with either glucose or ethanol and accumulates elevated pools of leucine; this phenotype is alleviated by the addition of leucine. Transformation of the leu4Δ mutant with a centromeric plasmid carrying LEU4 restored the wild-type phenotype. Bimolecular fluorescent complementation analysis showed that Leu4-Leu9 heterodimeric isozymes are formed in vivo. Purification and kinetic analysis showed that the hetero-oligomeric isozyme has a distinct leucine sensitivity behavior. Determination of α-IPMS activity in ethanol-grown cultures showed that α-IPM biosynthesis and growth under these respiratory conditions depend on the feedback-sensitive Leu4 homodimer. We conclude that retention and further diversification of two yeast α-IPMSs have resulted in a specific regulatory system that controls the leucine-α-IPM biosynthetic pathway by selective feedback sensitivity of homomeric and heterodimeric isoforms.

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来源期刊
Eukaryotic Cell
Eukaryotic Cell 生物-微生物学
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期刊介绍: Eukaryotic Cell (EC) focuses on eukaryotic microbiology and presents reports of basic research on simple eukaryotic microorganisms, such as yeasts, fungi, algae, protozoa, and social amoebae. The journal also covers viruses of these organisms and their organelles and their interactions with other living systems, where the focus is on the eukaryotic cell. Topics include: - Basic biology - Molecular and cellular biology - Mechanisms, and control, of developmental pathways - Structure and form inherent in basic biological processes - Cellular architecture - Metabolic physiology - Comparative genomics, biochemistry, and evolution - Population dynamics - Ecology
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