Kajimura Junko, Rahman Arifur, Hsu James, Evans Matthew R, Gardner Kevin H, Rick Paul D
Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814-4799.
J Bacteriol. 2006 Nov;188(21):7542-50. doi: 10.1128/JB.00783-06. Epub 2006 Aug 25.
The carbohydrate component of the enterobacterial common antigen (ECA) of Escherichia coli K-12 occurs primarily as a water-soluble cyclic polysaccharide located in the periplasm (ECA(CYC)) and as a phosphoglyceride-linked linear polysaccharide located on the cell surface (ECA(PG)). The polysaccharides of both forms are comprised of the amino sugars N-acetyl-D-glucosamine (GlcNAc), N-acetyl-D-mannosaminuronic acid (ManNAcA), and 4-acetamido-4,6-dideoxy-D-galactose (Fuc4NAc). These amino sugars are linked to one another to form trisaccharide repeat units with the structure -->3-alpha-D-Fuc4NAc-(1-->4)-beta-D-ManNAcA-(1-->4)-alpha-D-GlcNAc-(1-->. The hydroxyl group in the 6 position of the GlcNAc residues of both ECA(CYC) and ECA(PG) are nonstoichiometrically esterified with acetyl groups. Random transposon insertion mutagenesis of E. coli K-12 resulted in the generation of a mutant defective in the incorporation of O-acetyl groups into both ECA(CYC) and ECA(PG). This defect was found to be due to an insertion of the transposon into the yiaH locus, a putative gene of unknown function located at 80.26 min on the E. coli chromosomal map. Bioinformatic analyses of the predicted yiaH gene product indicate that it is an integral inner membrane protein that is a member of an acyltransferase family of enzymes found in a wide variety of organisms. The results of biochemical and genetic experiments presented here strongly support the conclusion that yiaH encodes the O-acetyltransferase responsible for the incorporation of O-acetyl groups into both ECA(CYC) and ECA(PG). Accordingly, we propose that this gene be designated wecH.
大肠杆菌K-12的肠杆菌共同抗原(ECA)的碳水化合物成分主要以位于周质中的水溶性环状多糖(ECA(CYC))和位于细胞表面的磷酸甘油酯连接的线性多糖(ECA(PG))形式存在。两种形式的多糖均由氨基糖N-乙酰-D-葡萄糖胺(GlcNAc)、N-乙酰-D-甘露糖胺糖醛酸(ManNAcA)和4-乙酰氨基-4,6-二脱氧-D-半乳糖(Fuc4NAc)组成。这些氨基糖相互连接形成具有结构-->3-α-D-Fuc4NAc-(1-->4)-β-D-ManNAcA-(1-->4)-α-D-GlcNAc-(1-->的三糖重复单元。ECA(CYC)和ECA(PG)的GlcNAc残基6位上的羟基非化学计量地被乙酰基酯化。大肠杆菌K-12的随机转座子插入诱变导致产生了一个在将O-乙酰基掺入ECA(CYC)和ECA(PG)方面有缺陷的突变体。发现该缺陷是由于转座子插入到yiaH基因座中,yiaH是位于大肠杆菌染色体图谱80.26分钟处的一个功能未知的推定基因。对预测的yiaH基因产物的生物信息学分析表明,它是一种整合内膜蛋白,是在多种生物体中发现的酰基转移酶家族的成员。本文给出的生化和遗传实验结果有力地支持了yiaH编码负责将O-乙酰基掺入ECA(CYC)和ECA(PG)的O-乙酰基转移酶这一结论。因此,我们建议将该基因命名为wecH。