The lipoprotein DolP affects cell separation in Escherichia coli, but not as an upstream regulator of NlpD.

The lipoprotein DolP affects cell separation in Escherichia coli, but not as an upstream regulator of NlpD.
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脂蛋白 DolP 影响大肠杆菌中的细胞分离,但不是 NlpD 的上游调节因子。

DOI:
10.1099/mic.0.001197
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发表时间:
2022
期刊:
Microbiology (Reading, England)
影响因子:
--
通讯作者:
Boelter G
Boelter G
中科院分区:
--
文献类型:
--
作者:
Boelter G

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细菌酰胺酶对于分裂附属子细胞的共有包膜以允许细胞分离是必需的。它们的活性需要精确控制以防止细胞裂解。在大肠杆菌中,酰胺酶活性受三个调节蛋白NlpD、EnvC和ActS控制。然而,最近的研究将外膜脂蛋白DolP(以前的YraP)作为NlpD的潜在上游调节因子联系起来。在这项研究中,我们进一步详细探讨了这一联系。令我们惊讶的是,DolP在体外不调节酰胺酶活性,并且在下拉和MST(MicroScale Thermophoresis)测定中不能与NlpD相互作用。其次,我们排除了在一定的包络应力范围内Δ dolP表观模仿Δ nlpD的假设。然而,对酰胺酶(AmiA,AmiB,AmiC)和酰胺酶调节因子(DolP)的双缺失突变体的形态学分析表明,ΔamiAΔ dolP和ΔenvCΔ dolP突变体与其亲本菌株相比显示出更长的链长,表明DolP在细胞分裂中的作用。总之,我们提出的证据表明,DolP不影响NlpD功能在体外,这意味着DolP不是一个上游调节NlpD。然而,DolP可能通过直接与AmiA或AmiC相互作用或通过尚未发现的机制影响子细胞分离。
Bacterial amidases are essential to split the shared envelope of adjunct daughter cells to allow cell separation. Their activity needs to be precisely controlled to prevent cell lysis. InEscherichia coli,amidase activity is controlled by three regulatory proteins NlpD, EnvC and ActS. However, recent studies linked the outer membrane lipoprotein DolP (formerly YraP) as a potential upstream regulator of NlpD. In this study we explored this link in further detail. To our surprise DolP did not modulate amidase activityin vitroand was unable to interact with NlpD in pull-down and MST (MicroScale Thermophoresis) assays. Next, we excluded the hypothesis that ΔdolPphenocopied ΔnlpDin a range of envelope stresses. However, morphological analysis of double deletion mutants of amidases (AmiA, AmiB AmiC) and amidase regulators withdolPrevealed that ΔamiAΔdolPand ΔenvCΔdolPmutants display longer chain length compared to their parental strains indicating a role for DolP in cell division. Overall, we present evidence that DolP does not affect NlpD functionin vitro, implying that DolP is not an upstream regulator of NlpD. However, DolP may impact daughter cell separation by interacting directly with AmiA or AmiC, or by a yet undiscovered mechanism.
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