Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Genome Biol. 2011 Oct 12;12(10):R99. doi: 10.1186/gb-2011-12-10-r99.
Two component regulatory systems are the primary form of signal transduction in bacteria. Although genomic binding sites have been determined for several eukaryotic and bacterial transcription factors, comprehensive identification of gene targets of two component response regulators remains challenging due to the lack of knowledge of the signals required for their activation. We focused our study on Desulfovibrio vulgaris Hildenborough, a sulfate reducing bacterium that encodes unusually diverse and largely uncharacterized two component signal transduction systems.
We report the first systematic mapping of the genes regulated by all transcriptionally acting response regulators in a single bacterium. Our results enabled functional predictions for several response regulators and include key processes of carbon, nitrogen and energy metabolism, cell motility and biofilm formation, and responses to stresses such as nitrite, low potassium and phosphate starvation. Our study also led to the prediction of new genes and regulatory networks, which found corroboration in a compendium of transcriptome data available for D. vulgaris. For several regulators we predicted and experimentally verified the binding site motifs, most of which were discovered as part of this study.
The gene targets identified for the response regulators allowed strong functional predictions to be made for the corresponding two component systems. By tracking the D. vulgaris regulators and their motifs outside the Desulfovibrio spp. we provide testable hypotheses regarding the functions of orthologous regulators in other organisms. The in vitro array based method optimized here is generally applicable for the study of such systems in all organisms.
双组分调控系统是细菌中信号转导的主要形式。尽管已经确定了几种真核和细菌转录因子的基因组结合位点,但由于缺乏对其激活所需信号的了解,全面鉴定双组分响应调节剂的基因靶标仍然具有挑战性。我们专注于研究脱硫弧菌(Desulfovibrio vulgaris Hildenborough),这是一种硫酸盐还原菌,其编码的双组分信号转导系统非常多样化,但大部分尚未得到表征。
我们首次系统地绘制了单个细菌中所有转录活性响应调节剂调节的基因图谱。我们的研究结果为几个响应调节剂提供了功能预测,包括碳、氮和能量代谢、细胞运动和生物膜形成以及对亚硝酸盐、低钾和磷酸盐饥饿等应激的反应等关键过程。我们的研究还预测了新的基因和调控网络,这些预测在可用于脱硫弧菌的转录组数据集的综合分析中得到了证实。对于几个我们预测并通过实验验证的调节剂,我们发现了部分结合基序,其中大多数是在这项研究中发现的。
响应调节剂的靶基因允许对相应的双组分系统进行强有力的功能预测。通过跟踪脱硫弧菌中的调节剂及其基序,我们为其他生物体中同源调节剂的功能提供了可测试的假设。这里优化的基于体外阵列的方法通常适用于所有生物体中此类系统的研究。