Functional analysis of selected sRNAs potentially involved in nitrogen and / or general stress response in the archaeon Methanosarcina mazei Gö1

对古细菌 Methanosarcina mazei Gö1 中可能参与氮和/或一般应激反应的选定 sRNA 进行功能分析

• 批准号: 38724498
• 负责人: Professorin Dr. Ruth Anne Schmitz-Streit
• 金额: --
• 依托单位: Institut für Allgemeine Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/38724498?language=en
• 关键词: Functional; analysis; selected; sRNAs; potentially

In previous work we identified 248 regulatory RNA (sRNA) candidates in Methanosarcina mazei, including sRNAs antisense to 5’ or 3’ regions of mRNAs (asRNAs), and sRNAs containing small ORFs of ≤ 30 amino acids (spRNAs). The strong conservation in Methanosarcinales and the high number of differentially transcribed sRNAs in response to nitrogen (N) argue for a prominent role of sRNAs in the N-stress response in M. mazei. We now aim to (i) study the regulatory role and mode of action of selected verified sRNAs potentially involved in N-regulation (e.g. sRNA154), (ii) elucidate the role of AmtR as a potential RNA binding protein and verify its proposed role in N-stress response, (iii) characterize the small ORFs identified in sRNAs and elucidate the potential dual function of the respective transcripts as mRNA and sRNAs, and (iv) study the proposed relationship of transposition events in M. mazei and N-availability by a post-transcriptional asRNA-mediated control of transposase expression. In order to achieve those major goals, several novel methods will be established for M. mazei including a tetracycline-inducible promoter for pulse-expression of sRNAs and fusing various RNA-tags to sRNAs to identify interacting partners. Overall, the proposed project will allow to unravel the role of sRNAs in the N-regulon of M. mazei and to achieve a deeper insight regarding the proposed transposition events in response to N availability in M. mazei.

在前期的工作中，我们在马氏甲烷八叠球菌中鉴定了248种调控RNA（sRNA），包括mRNA 5'或3'区的反义sRNA（asRNA）和含有≤ 30个氨基酸的小ORF的sRNA（spRNA）。甲烷藻目中的高度保守性和大量响应于氮（N）的差异转录sRNA证明了sRNA在甲烷藻的N胁迫响应中的突出作用。马泽我们现在的目标是（i）研究选择的验证sRNAs的调节作用和作用模式，这些sRNAs可能参与N-调节（例如sRNA 154），（ii）阐明AmtR作为潜在的RNA结合蛋白的作用并验证其在N-应激反应中的拟议作用，（iii）表征sRNA中鉴定的小ORF并阐明各自转录物作为mRNA和sRNA的潜在双重功能，（iv）研究M.通过转录后asRNA介导的转座酶表达的控制来控制Mazei和N-可用性。为了实现这些主要目标，将建立几个新的方法，为M。包括四环素诱导型启动子，用于sRNA的脉冲表达，并将各种RNA标签融合到sRNA以鉴定相互作用伴侣。总的来说，拟议的项目将允许解开sRNA在M的N-调节子中的作用。Mazei，并实现了更深的洞察力，关于建议的换位事件，以响应N的可用性在M。马泽