Cardiff School of Pharmacy and Pharmaceutical Science, Cardiff University , Cardiff, U.K. CF10 3NB.
Department of Biomolecular Sciences and Biotechnology, University of Milan , 20122 Milan, Italy.
Langmuir. 2016 Aug 9;32(31):7965-74. doi: 10.1021/acs.langmuir.6b02342. Epub 2016 Aug 1.
Curli are bacterial appendages involved in the adhesion of cells to surfaces; their synthesis is regulated by many genes such as csgD and ompR. The expression of the two curli subunits (CsgA and CsgB) in Escherichia coli (E. coli) is regulated by CsgD; at the same time, csgD transcription is under the control of OmpR. Therefore, both genes are involved in the control of curli production. In this work, we elucidated the role of these genes in the nanomechanical and adhesive properties of E. coli MG1655 (a laboratory strain not expressing significant amount of curli) and its curli-producing mutants overexpressing OmpR and CsgD, employing atomic force microscopy (AFM). Nanomechanical analysis revealed that the expression of these genes gave origin to cells with a lower Young's modulus (E) and turgidity (P0), whereas the adhesion forces were unaffected when genes involved in curli formation were expressed. AFM was also employed to study the primary structure of the curli expressed through the freely jointed chain (FJC) model for polymers. CsgD increased the number of curli on the surface more than OmpR did, and the overexpression of both genes did not result in a greater number of curli. Neither of the genes had an impact on the structure (total length of the polymer and number and length of Kuhn segments) of the curli. Our results further suggest that, despite the widely assumed role of curli in cell adhesion, cell adhesion force is also dictated by surface properties because no relation between the number of curli expressed on the surface and cell adhesion was found.
菌毛是一种细菌附属物,参与细胞与表面的黏附;其合成受许多基因的调控,如 csgD 和 ompR。大肠杆菌(Escherichia coli)中两种卷曲菌附属物(CsgA 和 CsgB)的表达受 CsgD 调控;同时,csgD 转录受 OmpR 控制。因此,这两个基因都参与卷曲菌产生的调控。在这项工作中,我们使用原子力显微镜(AFM)阐明了这些基因在大肠杆菌 MG1655(一种不表达大量卷曲菌的实验室菌株)及其过表达 OmpR 和 CsgD 的卷曲菌产生突变体的纳米力学和黏附特性中的作用。纳米力学分析表明,这些基因的表达导致细胞具有更低的杨氏模量(E)和膨压(P0),而卷曲菌形成相关基因的表达并不影响黏附力。AFM 还用于通过聚合物的自由连接链(FJC)模型研究表达的卷曲菌的初级结构。CsgD 比 OmpR 更能增加表面上的卷曲菌数量,而这两个基因的过表达并不会导致更多的卷曲菌。这两个基因都不会影响卷曲菌的结构(聚合物的总长度、Kuhn 片段的数量和长度)。我们的研究结果进一步表明,尽管卷曲菌在细胞黏附中具有广泛的作用,但细胞黏附力也取决于表面性质,因为在表面上表达的卷曲菌数量与细胞黏附之间没有关系。