Sorbonne Université, CNRS, Integrative Biology of Marine Models (LBI2M), Station Biologique de Roscoff (SBR), 29680 Roscoff, France.
Université Paris-Saclay, INRAE, UVSQ, VIM, 78350, Jouy-en-Josas, France.
Nucleic Acids Res. 2020 Aug 20;48(14):7786-7800. doi: 10.1093/nar/gkaa533.
Marine flavobacteria possess dedicated Polysaccharide Utilization Loci (PULs) enabling efficient degradation of a variety of algal polysaccharides. The expression of these PULs is tightly controlled by the presence of the substrate, yet details on the regulatory mechanisms are still lacking. The marine flavobacterium Zobellia galactanivorans DsijT digests many algal polysaccharides, including alginate from brown algae. Its complex Alginate Utilization System (AUS) comprises a PUL and several other loci. Here, we showed that the expression of the AUS is strongly and rapidly (<30 min) induced upon addition of alginate, leading to biphasic substrate utilization. Polymeric alginate is first degraded into smaller oligosaccharides that accumulate in the extracellular medium before being assimilated. We found that AusR, a GntR family protein encoded within the PUL, regulates alginate catabolism by repressing the transcription of most AUS genes. Based on our genetic, genomic, transcriptomic and biochemical results, we propose the first model of regulation for a PUL in marine bacteria. AusR binds to promoters of AUS genes via single, double or triple copies of operator. Upon addition of alginate, secreted enzymes expressed at a basal level catalyze the initial breakdown of the polymer. Metabolic intermediates produced during degradation act as effectors of AusR and inhibit the formation of AusR/DNA complexes, thus lifting transcriptional repression.
海洋黄杆菌具有专门的多糖利用基因座 (PUL),能够有效地降解多种藻类多糖。这些 PUL 的表达受底物的存在严格调控,但调控机制的细节仍不清楚。海洋黄杆菌 Zobellia galactanivorans DsijT 可消化多种藻类多糖,包括褐藻中的褐藻酸盐。其复杂的褐藻酸盐利用系统 (AUS) 由一个 PUL 和其他几个基因座组成。在这里,我们发现,褐藻酸盐的添加会强烈且迅速(<30 分钟)诱导 AUS 的表达,导致双相底物利用。聚合的褐藻酸盐首先被降解成较小的寡糖,在被吸收之前积累在细胞外介质中。我们发现,AUS 中的 GntR 家族蛋白编码产物 AusR 通过抑制大多数 AUS 基因的转录来调节褐藻酸盐的分解代谢。基于我们的遗传、基因组、转录组和生化结果,我们提出了海洋细菌中 PUL 调控的第一个模型。AusR 通过单个、双个或三个拷贝的操纵子结合到 AUS 基因的启动子上。褐藻酸盐添加后,基础水平表达的分泌酶催化聚合物的初始分解。在降解过程中产生的代谢中间产物作为 AusR 的效应物,抑制 AusR/DNA 复合物的形成,从而解除转录抑制。