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鉴定和表征……中的酵母SNF1激酶同源物

Identification and characterization of yeast SNF1 kinase homologs in .

作者信息

Shoeran Gaurav, Anand Namrata, Kaur Upninder, Goyal Kapil, Sehgal Rakesh

机构信息

Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

Department of Virology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

出版信息

Front Mol Biosci. 2025 Mar 24;12:1567703. doi: 10.3389/fmolb.2025.1567703. eCollection 2025.

Abstract

BACKGROUND

Sucrose Non Fermenting1 (SNF1) constitutes a family of protein kinases conserved in eukaryotes, plants, and fungi. SNF1 has been known to play a crucial role in stress adaptation and metabolism, enabling organisms to respond to changing environmental conditions. Initially identified in yeast, SNF1 is essential for shifting from the primary carbon source, glucose, to secondary carbon sources like sucrose. Homologs of this protein family were identified in , a protozoan parasite and we aimed to determine their role in this parasite.

METHODS

In the present study, we identified the putative homologs of SNF1 kinase in and knock out strains were prepared using the CRISPR-Cas9 knock-out strategy. The developed strains were evaluated for their growth, characteristics, protein expression and ultra structural changes and virulence in a mouse model.

RESULTS

One of the strain named N2, was found to be completely avirulent and showed limited growth, lack of glycosomes and had a fewer mitochondria with deformed cristae. The N2 strain failed to produce infection in mice when compared to WT mice. Proteome analysis revealed an increase in ribosomal proteins in the N2 strain, highlighting the role of ribosomes in stress adaptation.

CONCLUSION

The essentiality of this gene for developing infections in mice underscores its potential in the development of future antileishmanial therapies and live attenuated strains.

摘要

背景

蔗糖非发酵1(SNF1)构成了一个在真核生物、植物和真菌中保守的蛋白激酶家族。已知SNF1在应激适应和代谢中起关键作用,使生物体能够应对不断变化的环境条件。SNF1最初在酵母中被鉴定出来,对于从主要碳源葡萄糖向蔗糖等次要碳源的转变至关重要。在原生动物寄生虫利什曼原虫中鉴定出了这个蛋白家族的同源物,我们旨在确定它们在这种寄生虫中的作用。

方法

在本研究中,我们在利什曼原虫中鉴定了SNF1激酶的假定同源物,并使用CRISPR-Cas9敲除策略制备了敲除菌株。对所构建的菌株进行了生长、特征、蛋白表达、超微结构变化以及在小鼠模型中的毒力评估。

结果

发现其中一个名为N2的菌株完全无毒,生长受限,缺乏糖体,线粒体数量较少且嵴变形。与野生型小鼠相比,N2菌株在小鼠中未能引发感染。蛋白质组分析显示N2菌株中核糖体蛋白增加,突出了核糖体在应激适应中的作用。

结论

该基因对于在小鼠中引发感染的必要性强调了其在未来抗利什曼病治疗和减毒活菌株开发中的潜力。

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