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蛋白激酶Kic1影响酿酒酵母细胞壁中1,6-β-葡聚糖的水平。

The protein kinase Kic1 affects 1,6-beta-glucan levels in the cell wall of Saccharomyces cerevisiae.

作者信息

Vink Edwin, Vossen Jack H, Ram Arthur F J, van den Ende Herman, Brekelmans Stephan, de Nobel Hans, Klis Frans M

机构信息

Swammerdam Institute for Life Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018 WV, Amsterdam, The Netherlands1.

出版信息

Microbiology (Reading). 2002 Dec;148(Pt 12):4035-4048. doi: 10.1099/00221287-148-12-4035.

Abstract

KIC1 encodes a PAK kinase that is involved in morphogenesis and cell integrity. Both over- and underexpressing conditions of KIC1 affected cell wall composition. Kic1-deficient cells were hypersensitive to the cell wall perturbing agent calcofluor white and had less 1,6-beta-glucan. When Kic1-deficient cells were crossed with various kre mutants, which also have less 1,6-beta-glucan in their wall, the double mutants displayed synthetic growth defects. However, when crossed with the 1,3-beta-glucan-deficient strain fks1delta, no synthetic growth defect was observed, supporting a specific role for KIC1 in regulating 1,6-beta-glucan levels. Kic1-deficient cells also became highly resistant to the cell wall-degrading enzyme mixture Zymolyase, and exhibited higher transcript levels of the cell wall protein-encoding genes CWP2 and SED1. Conversely, overexpression of KIC1 resulted in increased sensitivity to Zymolyase and in a higher level of 1,6-beta-glucan. Multicopy suppressor analysis of a Kic1-deficient strain identified RHO3. Consistent with this, expression levels of RHO3 correlated with 1,6-beta-glucan levels in the cell wall. Interestingly, expression levels of KIC1 and the MAP kinase kinase PBS2 had opposite effects on Zymolyase sensitivity of the cells and on cell wall 1,6-beta-glucan levels in the wall. It is proposed that Kic1 affects cell wall construction in multiple ways and in particular in regulating 1,6-beta-glucan levels in the wall.

摘要

KIC1编码一种参与形态发生和细胞完整性的PAK激酶。KIC1的过表达和低表达条件均会影响细胞壁组成。Kic1缺陷型细胞对细胞壁干扰剂荧光增白剂高度敏感,且1,6-β-葡聚糖含量较低。当Kic1缺陷型细胞与各种kre突变体杂交时(这些突变体细胞壁中的1,6-β-葡聚糖含量也较低),双突变体表现出合成生长缺陷。然而,当与1,3-β-葡聚糖缺陷型菌株fks1δ杂交时,未观察到合成生长缺陷,这支持了KIC1在调节1,6-β-葡聚糖水平方面的特定作用。Kic1缺陷型细胞对细胞壁降解酶混合物溶壁酶也具有高度抗性,并表现出细胞壁蛋白编码基因CWP2和SED1的转录水平升高。相反,KIC1的过表达导致对溶壁酶的敏感性增加以及1,6-β-葡聚糖水平升高。对Kic1缺陷型菌株的多拷贝抑制子分析鉴定出RHO3。与此一致,RHO3的表达水平与细胞壁中1,6-β-葡聚糖水平相关。有趣的是,KIC1和丝裂原活化蛋白激酶激酶PBS2的表达水平对细胞对溶壁酶的敏感性以及细胞壁中1,6-β-葡聚糖水平具有相反的影响。有人提出,Kic1以多种方式影响细胞壁构建,特别是在调节细胞壁中1,6-β-葡聚糖水平方面。

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