Identification and phenotypic characterization of the cell-division protein CdpA
Identification and phenotypic characterization of the cell-division protein CdpA
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DOI：
10.1016/j.gene.2004.08.004
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发表时间：
2004-11-10
期刊：
影响因子：
3.5
通讯作者：
Klaenhammer, TR
中科院分区：
文献类型：
--
作者：
Altermann, E;Buck, BL;Klaenhammer, TR
对嗜酸乳杆菌NCFM的早期草案阶段基因组的自动计算机注释的分析揭示了先前发现的S层基因slpA和与S层蛋白具有弱相似性的另外的部分ORF。对整个基因进行测序，揭示了一个1799 bp的基因，编码599个氨基酸，计算分子量为64.8 kDa。在5 '区附近不能确定转录或翻译信号。然而，在该基因的直接下游鉴定出一个强的假定终止子，其自由能为-16.84 kcal/mol。一个PSI-Blast分析显示相似的S-层蛋白，细胞壁相关蛋白酶和己糖基转移酶的成员。计算无根系统发育树与其他的S-层蛋白质和蛋白酶的例子，分别从这两个群体中推出的蛋白质。L的一个衍生物。嗜酸乳杆菌NCFM通过靶向整合到基因中构建。与野生型相比，突变体细胞壁的非共价连接蛋白的SDS-PAGE分析揭示了细胞表面蛋白的损失。突变体的表型分析显示细胞形态发生了显着变化，改变了对各种环境应力的反应，并降低了细胞粘附力。基于计算机模拟和功能分析，我们确定该蛋白质在生长和细胞-细胞分离过程中的细胞壁加工中起作用，并将该基因命名为细胞分裂蛋白cdpA。(C)2004 Elsevier B. V.保留所有权利。
Analysis of the automated computer annotation of the early draft phase genome of Lactobacillus acidophilus NCFM revealed the previously discovered S-layer gene slpA and an additional partial ORF with weak similarities to S-layer proteins. The entire gene was sequenced to reveal a 1799-bp gene coding for 599 amino acids with a calculated molecular mass of 64.8 kDa. No transcription or translation signals could be determined in close proximity to the 5'-region. However, a strong putative terminator with a free energy of -16.84 kcal/mol was identified directly downstream of the gene. A PSI-Blast analysis showed similarities to members of S-layer proteins, cell-wall associated proteinases and hexosyl-transferases. Calculation of an unrooted phylogenetic tree with other examples of S-layer proteins and proteinases placed the deduced protein separately from both groups. A derivative of L. acidophilus NCFM was constructed by targeted integration into the gene. SDS-PAGE analysis of non-covalently linked proteins of the cell wall of the mutant, compared to the wild type, revealed the loss of a cell-surface protein. Phenotypic analyses of the mutant revealed significant changes in cell morphology, altered responses to various environmental stresses, and lowered cell adhesion. Based on the in silico and functional analyses, we ascertained that this protein plays a role in cell-wall processing during the growth and cell-cell separation and designated the gene as cell-division protein, cdpA. (C) 2004 Elsevier B.V. All rights reserved.