Department of Microbiology and Immunology, University of California at San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
Nucleic Acids Res. 2012 Apr;40(7):2907-24. doi: 10.1093/nar/gkr1190. Epub 2011 Dec 8.
Predicting the location and strength of promoters from genomic sequence requires accurate sequenced-based promoter models. We present the first model of a full-length bacterial promoter, encompassing both upstream sequences (UP-elements) and core promoter modules, based on a set of 60 promoters dependent on σ(E), an alternative ECF-type σ factor. UP-element contribution, best described by the length and frequency of A- and T-tracts, in combination with a PWM-based core promoter model, accurately predicted promoter strength both in vivo and in vitro. This model also distinguished active from weak/inactive promoters. Systematic examination of promoter strength as a function of RNA polymerase (RNAP) concentration revealed that UP-element contribution varied with RNAP availability and that the σ(E) regulon is comprised of two promoter types, one of which is active only at high concentrations of RNAP. Distinct promoter types may be a general mechanism for increasing the regulatory capacity of the ECF group of alternative σ's. Our findings provide important insights into the sequence requirements for the strength and function of full-length promoters and establish guidelines for promoter prediction and for forward engineering promoters of specific strengths.
从基因组序列预测启动子的位置和强度需要准确的基于测序的启动子模型。我们提出了第一个完整细菌启动子模型,该模型基于一组依赖于σ(E)的 60 个启动子,涵盖了上游序列(UP-elements)和核心启动子模块。基于长度和 A 和 T 链频率的 UP 元件贡献,结合基于 PWM 的核心启动子模型,准确预测了体内和体外启动子的强度。该模型还区分了活性启动子和弱/非活性启动子。对启动子强度作为 RNA 聚合酶 (RNAP) 浓度的函数的系统研究表明,UP 元件的贡献随 RNAP 的可用性而变化,并且σ(E)调控子由两种启动子类型组成,其中一种仅在 RNAP 的高浓度下才具有活性。不同的启动子类型可能是增加 ECF 组替代σ因子调控能力的一般机制。我们的研究结果为全长启动子的强度和功能的序列要求提供了重要的见解,并为启动子预测和特定强度的启动子正向工程提供了指导方针。