Venderbure C, Chastanet A, Boudsocq F, Sommer S, Bailone A
Institut Curie, Centre Universitaire, F-91405 Orsay, France.
J Bacteriol. 1999 Feb;181(4):1249-55. doi: 10.1128/JB.181.4.1249-1255.1999.
By its functional interaction with a RecA polymer, the mutagenic UmuD'C complex possesses an antirecombination activity. We show here that MucA'B, a functional homolog of the UmuD'C complex, inhibits homologous recombination as well. In F- recipients expressing MucA'B from a Ptac promoter, Hfr x F- recombination decreased with increasing MucA'B concentrations down to 50-fold. In damage-induced pKM101-containing cells expressing MucA'B from the native promoter, recombination between a UV-damaged F lac plasmid and homologous chromosomal DNA decreased 10-fold. Overexpression of MucA'B together with UmuD'C resulted in a synergistic inhibition of recombination. RecA[UmuR] proteins, which are resistant to UmuD'C inhibition of recombination, are inhibited by MucA'B while promoting MucA'B-promoted mutagenesis efficiently. The data suggest that MucA'B and UmuD'C contact a RecA polymer at distinct sites. The MucA'B complex was more active than UmuD'C in promoting UV mutagenesis, yet it did not inhibit recombination more than UmuD'C does. The enhanced mutagenic potential of MucA'B may result from its inherent superior capacity to assist DNA polymerase in trans-lesion synthesis. In the course of this work, we found that the natural plasmid pKM101 expresses around 45,000 MucA and 13,000 MucB molecules per lexA(Def) cell devoid of LexA. These molecular Muc concentrations are far above those of the chromosomally encoded Umu counterparts. Plasmid pKM101 belongs to a family of broad-host-range conjugative plasmids. The elevated levels of the Muc proteins might be required for successful installation of pKM101-like plasmids into a variety of host cells.
通过与RecA聚合物的功能相互作用,诱变型UmuD'C复合物具有抗重组活性。我们在此表明,UmuD'C复合物的功能同源物MucA'B也抑制同源重组。在从Ptac启动子表达MucA'B的F-受体中,Hfr×F-重组随着MucA'B浓度增加至50倍而降低。在从天然启动子表达MucA'B的损伤诱导的含pKM101的细胞中,紫外线损伤的F lac质粒与同源染色体DNA之间的重组降低了10倍。MucA'B与UmuD'C一起过表达导致对重组的协同抑制。对UmuD'C重组抑制具有抗性的RecA[UmuR]蛋白被MucA'B抑制,同时有效地促进MucA'B促进的诱变。数据表明MucA'B和UmuD'C在不同位点接触RecA聚合物。MucA'B复合物在促进紫外线诱变方面比UmuD'C更具活性,但它对重组的抑制作用并不比UmuD'C更强。MucA'B诱变潜力的增强可能源于其在协助DNA聚合酶进行跨损伤合成方面固有的优越能力。在这项工作过程中,我们发现天然质粒pKM101在每个缺乏LexA的lexA(Def)细胞中表达约45,000个MucA分子和13,000个MucB分子。这些分子Muc浓度远高于染色体编码的Umu对应物。质粒pKM101属于广泛宿主范围的接合质粒家族。Muc蛋白的高水平可能是将pKM101样质粒成功导入各种宿主细胞所必需的。