Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Appl Environ Microbiol. 2013 Dec;79(23):7188-202. doi: 10.1128/AEM.02148-13. Epub 2013 Sep 13.
Cellulose fibrils play a role in attachment of Agrobacterium tumefaciens to its plant host. While the genes for cellulose biosynthesis in the bacterium have been identified, little is known concerning the regulation of the process. The signal molecule cyclic di-GMP (c-di-GMP) has been linked to the regulation of exopolysaccharide biosynthesis in many bacterial species, including A. tumefaciens. In this study, we identified two putative diguanylate cyclase genes, celR (atu1297) and atu1060, that influence production of cellulose in A. tumefaciens. Overexpression of either gene resulted in increased cellulose production, while deletion of celR, but not atu1060, resulted in decreased cellulose biosynthesis. celR overexpression also affected other phenotypes, including biofilm formation, formation of a polar adhesion structure, plant surface attachment, and virulence, suggesting that the gene plays a role in regulating these processes. Analysis of celR and Δcel mutants allowed differentiation between phenotypes associated with cellulose production, such as biofilm formation, and phenotypes probably resulting from c-di-GMP signaling, which include polar adhesion, attachment to plant tissue, and virulence. Phylogenetic comparisons suggest that species containing both celR and celA, which encodes the catalytic subunit of cellulose synthase, adapted the CelR protein to regulate cellulose production while those that lack celA use CelR, called PleD, to regulate specific processes associated with polar localization and cell division.
纤维素原纤维在根瘤农杆菌与其植物宿主附着中发挥作用。虽然该细菌中纤维素生物合成的基因已被鉴定,但该过程的调控知之甚少。信号分子环二鸟苷酸(c-di-GMP)已与许多细菌物种(包括根瘤农杆菌)中多糖生物合成的调控有关。在这项研究中,我们鉴定了两个假定的双鸟苷酸环化酶基因 celR(atu1297)和 atu1060,它们影响根瘤农杆菌中纤维素的产生。这两个基因的过表达都导致纤维素的产生增加,而 celR 的缺失,而不是 atu1060 的缺失,导致纤维素生物合成减少。celR 的过表达也影响了其他表型,包括生物膜形成、极性附着结构的形成、植物表面附着和毒性,表明该基因在调节这些过程中发挥作用。celR 和Δcel 突变体的分析允许区分与纤维素产生相关的表型,如生物膜形成,以及可能由 c-di-GMP 信号传导引起的表型,包括极性附着、与植物组织的附着和毒性。系统发育比较表明,同时含有 celR 和 celA 的物种,celA 编码纤维素合酶的催化亚基,使 CelR 蛋白适应调节纤维素的产生,而那些缺乏 celA 的物种则使用 CelR,称为 PleD,来调节与极性定位和细胞分裂相关的特定过程。