Schulz Annina, Hermann Lucas, Freibert Sven-Andreas, Bönig Tobias, Hoffmann Tamara, Riclea Ramona, Dickschat Jeroen S, Heider Johann, Bremer Erhard
Department of Biology, Laboratory for Microbiology, Philipps-University Marburg, Karl-von-Frisch-Str. 8, D-35043 Marburg, Germany.
Department of Medicine, Institute for Cytobiology and Cytopathology, Philipps-University Marburg, Robert-Koch Str. 6, D-35032 Marburg, Germany.
Environ Microbiol. 2017 Nov;19(11):4599-4619. doi: 10.1111/1462-2920.13924. Epub 2017 Oct 13.
Ectoine and hydroxyectoine are effective microbial osmostress protectants, but can also serve as versatile nutrients for bacteria. We have studied the genetic regulation of ectoine and hydroxyectoine import and catabolism in the marine Roseobacter species Ruegeria pomeroyi and identified three transcriptional regulators involved in these processes: the GabR/MocR-type repressor EnuR, the feast and famine-type regulator AsnC and the two-component system NtrYX. The corresponding genes are widely associated with ectoine and hydroxyectoine uptake and catabolic gene clusters (enuR, asnC), and with microorganisms predicted to consume ectoines (ntrYX). EnuR contains a covalently bound pyridoxal-5'-phosphate as a co-factor and the chemistry underlying the functioning of MocR/GabR-type regulators typically requires a system-specific low molecular mass effector molecule. Through ligand binding studies with purified EnuR, we identified N-(alpha)-L-acetyl-2,4-diaminobutyric acid and L-2,4-diaminobutyric acid as inducers for EnuR that are generated through ectoine catabolism. AsnC/Lrp-type proteins can wrap DNA into nucleosome-like structures, and we found that the asnC gene was essential for use of ectoines as nutrients. Furthermore, we discovered through transposon mutagenesis that the NtrYX two-component system is required for their catabolism. Database searches suggest that our findings have important ramifications for an understanding of the molecular biology of most microbial consumers of ectoines.
四氢嘧啶和羟基四氢嘧啶是有效的微生物渗透胁迫保护剂,但也可作为细菌的通用营养物质。我们研究了海洋玫瑰杆菌属的波氏鲁杰氏菌(Ruegeria pomeroyi)中四氢嘧啶和羟基四氢嘧啶的摄取及分解代谢的基因调控,并鉴定出参与这些过程的三个转录调节因子:GabR/MocR型阻遏蛋白EnuR、丰歉型调节因子AsnC和双组分系统NtrYX。相应的基因广泛与四氢嘧啶和羟基四氢嘧啶摄取及分解代谢基因簇(enuR、asnC)以及预测会消耗四氢嘧啶的微生物(ntrYX)相关。EnuR含有共价结合的磷酸吡哆醛作为辅因子,而MocR/GabR型调节因子发挥功能的化学机制通常需要一个系统特异性的低分子量效应分子。通过对纯化的EnuR进行配体结合研究,我们鉴定出N-(α)-L-乙酰-2,4-二氨基丁酸和L-2,4-二氨基丁酸是通过四氢嘧啶分解代谢产生的EnuR诱导剂。AsnC/Lrp型蛋白可将DNA包裹成核小体样结构,我们发现asnC基因对于将四氢嘧啶用作营养物质至关重要。此外,我们通过转座子诱变发现,NtrYX双组分系统是其分解代谢所必需的。数据库搜索表明,我们的发现对于理解大多数四氢嘧啶微生物消费者的分子生物学具有重要意义。