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整合生化、生物物理和转录组学数据以研究酵母细胞壁的结构和纳米力学特性

Integration of Biochemical, Biophysical and Transcriptomics Data for Investigating the Structural and Nanomechanical Properties of the Yeast Cell Wall.

作者信息

Schiavone Marion, Déjean Sébastien, Sieczkowski Nathalie, Castex Mathieu, Dague Etienne, François Jean M

机构信息

Laboratoire d'Ingénierie des Systèmes Biologiques et Procédés, Institut National des Sciences Appliquées de Toulouse, UPS, INP, Université de ToulouseToulouse, France.

Lallemand SASBlagnac, France.

出版信息

Front Microbiol. 2017 Sep 27;8:1806. doi: 10.3389/fmicb.2017.01806. eCollection 2017.

Abstract

The yeast cell is surrounded by a cell wall conferring protection and resistance to environmental conditions that can be harmful. Identify the molecular cues (genes) which shape the biochemical composition and the nanomechanical properties of the cell wall and the links between these two parameters represent a major issue in the understanding of the biogenesis and the molecular assembly of this essential cellular structure, which may have consequences in diverse biotechnological applications. We addressed this question in two ways. Firstly, we compared the biochemical and biophysical properties using atomic force microscopy (AFM) methods of 4 industrial strains with the laboratory sequenced strain BY4743 and used transcriptome data of these strains to infer biological hypothesis about differences of these properties between strains. This comparative approach showed a 4-6-fold higher hydrophobicity of industrial strains that was correlated to higher expression of genes encoding adhesin and adhesin-like proteins and not to their higher mannans content. The second approach was to employ a multivariate statistical analysis to identify highly correlated variables among biochemical, biophysical and genes expression data. Accordingly, we found a tight association between hydrophobicity and adhesion events that positively correlated with a set of 22 genes in which the main enriched GO function was the sterol metabolic process. We also identified a strong association of β-1,3-glucans with contour length that corresponds to the extension of mannans chains upon pulling the mannosyl units with the lectin-coated AFM tips. This association was positively correlated with a group of 27 genes in which the seripauperin multigene family was highly documented and negatively connected with a set of 23 genes whose main GO biological process was sulfur assimilation/cysteine biosynthetic process. On the other hand, the elasticity modulus was found weakly associated with levels of β-1,6-glucans, and this biophysical variable was positively correlated with a set of genes implicated in microtubules polymerization, tubulin folding and mitotic organization.

摘要

酵母细胞被一层细胞壁所包围,这层细胞壁赋予细胞保护作用,并使其能够抵御可能有害的环境条件。确定塑造细胞壁生化组成和纳米力学特性的分子线索(基因),以及这两个参数之间的联系,是理解这一重要细胞结构的生物发生和分子组装的一个主要问题,这可能会对多种生物技术应用产生影响。我们通过两种方式解决了这个问题。首先,我们使用原子力显微镜(AFM)方法比较了4种工业菌株与实验室测序菌株BY4743的生化和生物物理特性,并利用这些菌株的转录组数据推断关于菌株间这些特性差异的生物学假设。这种比较方法显示,工业菌株的疏水性高4至6倍,这与编码粘附素和粘附素样蛋白的基因的高表达相关,而与它们较高的甘露聚糖含量无关。第二种方法是采用多变量统计分析来确定生化、生物物理和基因表达数据之间高度相关的变量。因此,我们发现疏水性与粘附事件之间存在紧密关联,粘附事件与一组22个基因呈正相关,其中主要富集的GO功能是甾醇代谢过程。我们还确定了β-1,3-葡聚糖与轮廓长度之间的强关联,轮廓长度对应于用凝集素包被的AFM探针拉动甘露糖基单元时甘露聚糖链的延伸。这种关联与一组27个基因呈正相关,其中丝氨酸蛋白酶多基因家族有大量记录,并且与一组23个基因呈负相关,这组基因的主要GO生物学过程是硫同化/半胱氨酸生物合成过程。另一方面,发现弹性模量与β-1,6-葡聚糖水平的关联较弱,并且这个生物物理变量与一组涉及微管聚合、微管蛋白折叠和有丝分裂组织的基因呈正相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4a54/5649194/fc16e6f56258/fmicb-08-01806-g0001.jpg

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