Kloser A W, Laird M W, Misra R
Department of Microbiology, Arizona State University, Tempe 85287-2701, USA.
J Bacteriol. 1996 Sep;178(17):5138-43. doi: 10.1128/jb.178.17.5138-5143.1996.
A novel genetic scheme allowed us to isolate extragenic suppressor mutations that restored mutant OmpF assembly. One group of these mutations, termed asmB for assembly suppressor mutation B, permitted mutant OmpF assembly in a non-allele-specific manner. Genetic mapping analyses placed the asmB mutations at the 2-min region of the Escherichia coli K-12 chromosome. Further analyses revealed that the asmB mutations map within the envA (lpxC) gene, which encodes an enzyme needed for the synthesis of the lipid A moiety of lipopolysaccharide (LPS). Nucleotide sequence analysis showed that the asmB mutations caused a change from F-50 to S (F50S substitution) (asmB2 and asmB3) or a G210S substitution (asmB1) in EnvA. Cells bearing the asmB alleles displayed increased sensitivity to various hydrophobic compounds and detergents, suggesting an alteration within the outer membrane. Direct examination (of the LPS showed that its amounts were reduced by the asmB mutations, with asmB1 exerting a greater effect than asmB2 or asmB3. Thus, it appears that the asmB mutations achieve mutant OmpF assembly suppression by reducing LPS levels, which in turn may alter membrane fluidity.
一种新的遗传方案使我们能够分离出恢复突变型OmpF组装的基因外抑制突变。其中一组突变,称为组装抑制突变B(asmB),以非等位基因特异性方式允许突变型OmpF组装。遗传图谱分析将asmB突变定位在大肠杆菌K-12染色体的2分钟区域。进一步分析表明,asmB突变位于envA(lpxC)基因内,该基因编码脂多糖(LPS)脂质A部分合成所需的一种酶。核苷酸序列分析表明,asmB突变导致EnvA中F-50变为S(F50S替换)(asmB2和asmB3)或G210S替换(asmB1)。携带asmB等位基因的细胞对各种疏水化合物和去污剂的敏感性增加,表明外膜发生了改变。对LPS的直接检查表明,asmB突变使其含量降低,其中asmB1的影响大于asmB2或asmB3。因此,asmB突变似乎通过降低LPS水平来实现对突变型OmpF组装的抑制,这反过来可能会改变膜流动性。