Deciphering the role of the two conserved motifs of the ECF41 family σ factor in the autoregulation of its own promoter in Azospirillum brasilense Sp245

Ekta Pathak, A. Dubey, V. Singh, Rajeev Mishra, A. Tripathi
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Abstract

In Azospirillum brasilense, an extra‐cytoplasmic function σ factor (RpoE10) shows the characteristic 119 amino acid long C‐terminal extension found in ECF41‐type σ factors, which possesses three conserved motifs (WLPEP, DGGGR, and NPDKV), one in the linker region between the σ2 and σ4, and the other two in the SnoaL_2 domain of the C‐terminal extension. Here, we have described the role of the two conserved motifs in the SnoaL_2 domain of RpoE10 in the inhibition and activation of its activity, respectively. Truncation of the distal part of the C‐terminal sequence of the RpoE10 (including NPDKV but excluding the DGGGR motif) results in its promoter's activation suggesting autoregulation. Further truncation of the C‐terminal sequence up to its proximal part, including NPDKV and DGGGR motif, abolished promoter activation. Replacement of NPDKV motif with NAAAV in RpoE10 increased its ability to activate its promoter, whereas replacement of DGGGR motif led to reduced promoter activation. We have explored the dynamic modulation of σ2 ‐σ4 domains and the relevant molecular interactions mediated by the two conserved motifs of the SnoaL2 domain using molecular dynamics simulation. The analysis enabled us to explain that the NPDKV motif located distally in the C‐terminus negatively impacts transcriptional activation. In contrast, the DGGGR motif found proximally of the C‐terminal extension is required to activate RpoE10.
解读巴西氮螺旋菌Sp245中ECF41家族σ因子的两个保守基序在其自身启动子自动调控中的作用
在巴西偶氮螺旋菌中,胞质外功能σ因子(RpoE10)具有ECF41型σ因子特有的119个氨基酸长的C末端延伸,具有3个保守基序(WLPEP、DGGGR和NPDKV),一个位于σ2和σ4之间的连接区,另外两个位于C末端延伸的SnoaL_2结构域。在这里,我们分别描述了RpoE10的SnoaL_2结构域的两个保守基序在抑制和激活其活性中的作用。RpoE10 C末端序列的末端部分(包括NPDKV但不包括DGGGR基序)的截断导致其启动子的激活,提示自动调节。进一步截断C端序列直至其近端部分,包括NPDKV和DGGGR基序,取消启动子激活。用NAAAV替换RpoE10中的NPDKV基序增加了其激活启动子的能力,而替换DGGGR基序导致启动子激活降低。本文采用分子动力学模拟的方法,探讨了SnoaL2结构域的两个保守基序对σ2‐σ4结构域的动态调制和相关的分子相互作用。该分析使我们能够解释位于C端远端的NPDKV基序对转录激活产生负面影响。相比之下,在C末端延伸的近端发现的DGGGR基序是激活RpoE10所必需的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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