TY - JOUR
T1 - The lipoprotein DolP affects cell separation in Escherichia colbut not as an upstream regulator of NlpD
AU - Boelter, Gabriela
AU - Bryant, Jack A.
AU - Doherty, Hannah
AU - Wotherspoon, Peter
AU - Alodaini, Dema
AU - Ma, Xuyu
AU - Alao, Micheal B.
AU - Moynihan, Patrick J.
AU - Moradigaravand, Danesh
AU - Glinkowska, Monika
AU - Knowles, Timothy J.
AU - Henderson, Ian R.
AU - Banzhaf, Manuel
N1 - Generated from Scopus record by KAUST IRTS on 2023-02-15
PY - 2022/1/1
Y1 - 2022/1/1
N2 - 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.
AB - 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.
UR - https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.001197
UR - http://www.scopus.com/inward/record.url?scp=85130480027&partnerID=8YFLogxK
U2 - 10.1099/mic.0.001197
DO - 10.1099/mic.0.001197
M3 - Article
C2 - 35604759
SN - 1350-0872
VL - 168
JO - Microbiology (United Kingdom)
JF - Microbiology (United Kingdom)
IS - 5
ER -