MOLECULAR-CLONING OF CWP1 - A GENE ENCODING A SACCHAROMYCES-CEREVISIAE CELL-WALL PROTEIN SOLUBILIZED WITH RAROBACTER-FAECITABIDUS PROTEASE-I

被引:54
作者
SHIMOI, H
IIMURA, Y
OBATA, T
机构
[1] National Research Institute of Brewing, Kita-ku, Tokyo 114
关键词
CELL WALL PROTEIN; GENE CLONING; GLYCOSYLPHOSPHATIDYLINOSITOL-ANCHORED PROTEIN; RAROBACTER FAECITABIDUS PROTEASE I; SACCHAROMYCES CEREVISIAE;
D O I
10.1093/oxfordjournals.jbchem.a124907
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A yeast cell wall glycoprotein with a molecular weight of 40,000, named gp40, was solubilized from SDS-extracted cell wall of Snccharomyces cerevisiae by incubation with Rarobacter faecitabidus protease I, which is a yeast-lytic enzyme, Based on its amino acid sequence, we cloned and sequenced the gene encoding the precursor of gp40, named CWP1; cell wall protein gene, The DNA sequence of the CWP1 gene was identical to YKL443, an open reading frame identified in a genome sequencing program for yeast chromosome XI. This gene encoded a serine-rich protein of 239 amino acids with a molecular weight of 24,267. The presence of hydrophobic sequences in the N- and C-termini of the CWP1 protein suggests that it is secreted as a glycosylphosphatidylinositol-anchored protein and is subsequently integrated into the cell wall. Since a gene disruption experiment showed no growth defect, the CWP1 gene is not essential for growth. Mutant CWP1 protein deficient in the C-terminal hydrophobic sequence was secreted into the culture medium, not anchored to the cell wall, thereby indicating that this hydrophobic sequence plays a crucial role in anchoring to the cell wall. Homology between the CWP1 protein and TIP1 family of cold shock proteins suggests that they belong to a new family of cell wall proteins.
引用
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页码:302 / 311
页数:10
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