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

Lookup NU author(s): Manuel Banzhaf

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

© 2022 The Authors. 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. In Escherichia 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 activity in vitro and was unable to interact with NlpD in pull-down and MST (MicroScale Thermophoresis) assays. Next, we excluded the hypothesis that ΔdolP phenocopied ΔnlpD in a range of envelope stresses. However, morphological analysis of double deletion mutants of amidases (AmiA, AmiB AmiC) and amidase regulators with dolP revealed that ΔamiAΔdolP and ΔenvCΔdolP mutants 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 function in 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.


Publication metadata

Author(s): Boelter G, Bryant JA, Doherty H, Wotherspoon P, Alodaini D, Ma X, Alao MB, Moynihan PJ, Moradigaravand D, Glinkowska M, Knowles TJ, Henderson IR, Banzhaf M

Publication type: Article

Publication status: Published

Journal: Microbiology

Year: 2022

Volume: 168

Issue: 5

Online publication date: 23/05/2022

Acceptance date: 06/05/2022

Date deposited: 09/08/2023

ISSN (print): 1350-0872

ISSN (electronic): 1465-2080

Publisher: Microbiology Society

URL: https://doi.org/10.1099/mic.0.001197

DOI: 10.1099/mic.0.001197

PubMed id: 35604759


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Funding

Funder referenceFunder name
Narodowym Centrum Nauki
MR/V027204/1
UKRI
UMO-2017/27/B/NZ2/00747

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