Disruption of c-di-GMP Signaling Networks Unlocks Cryptic Expression of Secondary Metabolites during Biofilm Growth in Burkholderia pseudomallei.
Disruption of c-di-GMP Signaling Networks Unlocks Cryptic Expression of Secondary Metabolites during Biofilm Growth in Burkholderia pseudomallei.
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DOI：
10.1128/aem.02431-21
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发表时间：
2022-04-26
影响因子：
4.4
通讯作者：
Borlee, Bradley R.
中科院分区：
文献类型：
--
作者：
Borlee, Grace, I;Mangalea, Mihnea R.;Martin, Kevin H.;Plumley, Brooke A.;Golon, Samuel J.;Borlee, Bradley R.
伯克霍尔德氏菌生物膜生长过程中次生代谢产物的产生与调控。并没有得到很好的理解。为了了解更多关于生物膜生长过程中产生的隐蔽分子的关键作用和调控，我们破坏了类鼻疽伯克霍尔德氏菌（一种土传细菌saccharide和类鼻疽的病原体）中的c-di-GMP信号传导。我们对这些研究的方法结合了转录谱分析和基因缺失，靶向关键的c-di-GMP调控成分，以表征对温度变化的反应。c-di-GMP基因的突变分析和条件表达研究表明，当在不同温度下作为生物膜生长时，它们对表型如生物膜形成、菌落形态、运动性和次级代谢产物生物合成的表达的贡献。在不同温度下在Δ II 2523突变体背景中进行RNA-seq分析，该突变体背景响应于导致低生物膜和高生物膜形成表型的温度变化。差异调节的基因被观察到多糖的生物合成，分泌系统，和非核糖体肽和聚酮合酶（NRPS/PKS）集群响应温度的变化。亲本和Δ II 2523背景中生物合成基因簇（BGC）2、11、14（阿伐他汀）和15（马利抑菌素）的缺失突变也揭示了这些BGC除了抑制枯草芽孢杆菌和立枯丝核菌外，还对生物膜形成和菌落形态的贡献。我们的研究结果表明，II 2523影响基因的调控，有助于生物膜的形成和竞争。在不同的环境条件下的隐蔽BGC的表征将允许更好地理解在生物膜形成和微生物-微生物相互作用的背景下的次级代谢产物的作用。重要性类鼻疽伯克霍尔德菌是一种寄生在环境中的致病菌，在感染广泛的宿主期间转变为致病性生活方式。作为触发这种变化的刺激因素的环境线索在很大程度上是未知的。然而，已经确定的是，c-di-GMP（第二信号信使）的细胞水平控制从作为增殖细胞生长到作为生物膜生长的转换。破坏由c-di-GMP介导的信号传导允许更好地理解有助于该生物体的各种生活方式的表面相关和分泌分子的调节和贡献。B的基因组。类鼻疽杆菌还编码隐蔽的生物合成基因簇，该基因簇被预测编码可能有助于作为生物膜生长、适应和与其它生物体相互作用的小分子。更好地了解这些分子的调节对于了解这种多功能病原体如何改变其生活方式至关重要。
The regulation and production of secondary metabolites during biofilm growth of Burkholderia spp. is not well understood. To learn more about the crucial role and regulatory control of cryptic molecules produced during biofilm growth, we disrupted c-di-GMP signaling in Burkholderia pseudomallei, a soilborne bacterial saprophyte and the etiologic agent of melioidosis. Our approach to these studies combined transcriptional profiling with genetic deletions that targeted key c-di-GMP regulatory components to characterize responses to changes in temperature. Mutational analyses and conditional expression studies of c-di-GMP genes demonstrates their contribution to phenotypes such as biofilm formation, colony morphology, motility, and expression of secondary metabolite biosynthesis when grown as a biofilm at different temperatures. RNA-seq analysis was performed at various temperatures in a ΔII2523 mutant background that is responsive to temperature alterations resulting in hypobiofilm- and hyperbiofilm-forming phenotypes. Differential regulation of genes was observed for polysaccharide biosynthesis, secretion systems, and nonribosomal peptide and polyketide synthase (NRPS/PKS) clusters in response to temperature changes. Deletion mutations of biosynthetic gene clusters (BGCs) 2, 11, 14 (syrbactin), and 15 (malleipeptin) in parental and ΔII2523 backgrounds also reveal the contribution of these BGCs to biofilm formation and colony morphology in addition to inhibition of Bacillus subtilis and Rhizoctonia solani. Our findings suggest that II2523 impacts the regulation of genes that contribute to biofilm formation and competition. Characterization of cryptic BGCs under different environmental conditions will allow for a better understanding of the role of secondary metabolites in the context of biofilm formation and microbe-microbe interactions. IMPORTANCE Burkholderia pseudomallei is a saprophytic bacterium residing in the environment that switches to a pathogenic lifestyle during infection of a wide range of hosts. The environmental cues that serve as the stimulus to trigger this change are largely unknown. However, it is well established that the cellular level of c-di-GMP, a secondary signal messenger, controls the switch from growth as planktonic cells to growth as a biofilm. Disrupting the signaling mediated by c-di-GMP allows for a better understanding of the regulation and the contribution of the surface associated and secreted molecules that contribute to the various lifestyles of this organism. The genome of B. pseudomallei also encodes cryptic biosynthetic gene clusters predicted to encode small molecules that potentially contribute to growth as a biofilm, adaptation, and interactions with other organisms. A better understanding of the regulation of these molecules is crucial to understanding how this versatile pathogen alters its lifestyle.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
DOI：
10.1099/00207713-48-1-317
发表时间：
1998-01-01
期刊：
INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY
影响因子：
--
作者：
Brett, PJ;DeShazer, D;Woods, DE
通讯作者：
Woods, DE
影响因子：
4.4
作者：
Hall, Carina M.;Baker, Anthony L.;Wagner, David M.
通讯作者：
Wagner, David M.
影响因子：
11.8
作者：
Gee JE;Elrod MG;Gulvik CA;Haselow DT;Waters C;Liu L;Hoffmaster AR
通讯作者：
Hoffmaster AR