Cargando…

Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle

During cell division in Escherichia coli, the highly conserved tubulin homolog FtsZ polymerizes and assembles into a ring-like structure, called the Z-ring, at the site of septation. For recruitment to the membrane surface, FtsZ polymers directly interact with membrane-associated proteins, predomina...

Descripción completa

Detalles Bibliográficos
Autores principales: Morrison, Josiah J., Ferreira, Colby N., Siler, Evelyn M., Nelson, Katie, Trebino, Catherine E., Piraino, Benjamin, Camberg, Jodi L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213515/
https://www.ncbi.nlm.nih.gov/pubmed/37250038
http://dx.doi.org/10.3389/fmicb.2023.1171376
_version_ 1785047636354531328
author Morrison, Josiah J.
Ferreira, Colby N.
Siler, Evelyn M.
Nelson, Katie
Trebino, Catherine E.
Piraino, Benjamin
Camberg, Jodi L.
author_facet Morrison, Josiah J.
Ferreira, Colby N.
Siler, Evelyn M.
Nelson, Katie
Trebino, Catherine E.
Piraino, Benjamin
Camberg, Jodi L.
author_sort Morrison, Josiah J.
collection PubMed
description During cell division in Escherichia coli, the highly conserved tubulin homolog FtsZ polymerizes and assembles into a ring-like structure, called the Z-ring, at the site of septation. For recruitment to the membrane surface, FtsZ polymers directly interact with membrane-associated proteins, predominantly FtsA in E. coli. FtsA shares structural homology with actin and, like actin, hydrolyzes ATP. Yeast actin detects nucleotide occupancy through a sensor region adjacent to the nucleotide binding site and adopts distinct conformations in monomeric and filamentous actin. Bacterial actin homologs also display considerable conformational flexibility across different nucleotide-bound states and polymerize. Here, we show that several amino acid residues proximal to the nucleotide binding site in FtsA are critical for function in vitro and in vivo. Each of these residues are important for ATP hydrolysis, phospholipid (PL) binding, ATP-dependent vesicle remodeling, and recruitment to the divisome in vivo, to varying degrees. Notably, we observed that Ser 84 and Glu 14 are essential for ATP-dependent vesicle remodeling and magnesium-dependent membrane release of FtsA from vesicles in vitro, and these defects likely underlie the loss of function by FtsA(E14R) and FtsA(S84L) in vivo. Finally, we demonstrate that FtsA(A188V), which is associated with temperature-sensitive growth in vivo, is defective for rapid ATP hydrolysis and ATP-dependent remodeling of PL vesicles in vitro. Together, our results show that loss of nucleotide-dependent activities by FtsA, such as ATP hydrolysis, membrane binding and release, and, most importantly, ATP-dependent PL remodeling, lead to failed Z-ring assembly and division defects in cells.
format Online
Article
Text
id pubmed-10213515
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-102135152023-05-27 Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle Morrison, Josiah J. Ferreira, Colby N. Siler, Evelyn M. Nelson, Katie Trebino, Catherine E. Piraino, Benjamin Camberg, Jodi L. Front Microbiol Microbiology During cell division in Escherichia coli, the highly conserved tubulin homolog FtsZ polymerizes and assembles into a ring-like structure, called the Z-ring, at the site of septation. For recruitment to the membrane surface, FtsZ polymers directly interact with membrane-associated proteins, predominantly FtsA in E. coli. FtsA shares structural homology with actin and, like actin, hydrolyzes ATP. Yeast actin detects nucleotide occupancy through a sensor region adjacent to the nucleotide binding site and adopts distinct conformations in monomeric and filamentous actin. Bacterial actin homologs also display considerable conformational flexibility across different nucleotide-bound states and polymerize. Here, we show that several amino acid residues proximal to the nucleotide binding site in FtsA are critical for function in vitro and in vivo. Each of these residues are important for ATP hydrolysis, phospholipid (PL) binding, ATP-dependent vesicle remodeling, and recruitment to the divisome in vivo, to varying degrees. Notably, we observed that Ser 84 and Glu 14 are essential for ATP-dependent vesicle remodeling and magnesium-dependent membrane release of FtsA from vesicles in vitro, and these defects likely underlie the loss of function by FtsA(E14R) and FtsA(S84L) in vivo. Finally, we demonstrate that FtsA(A188V), which is associated with temperature-sensitive growth in vivo, is defective for rapid ATP hydrolysis and ATP-dependent remodeling of PL vesicles in vitro. Together, our results show that loss of nucleotide-dependent activities by FtsA, such as ATP hydrolysis, membrane binding and release, and, most importantly, ATP-dependent PL remodeling, lead to failed Z-ring assembly and division defects in cells. Frontiers Media S.A. 2023-05-12 /pmc/articles/PMC10213515/ /pubmed/37250038 http://dx.doi.org/10.3389/fmicb.2023.1171376 Text en Copyright © 2023 Morrison, Ferreira, Siler, Nelson, Trebino, Piraino and Camberg. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Morrison, Josiah J.
Ferreira, Colby N.
Siler, Evelyn M.
Nelson, Katie
Trebino, Catherine E.
Piraino, Benjamin
Camberg, Jodi L.
Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title_full Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title_fullStr Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title_full_unstemmed Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title_short Nucleotide-dependent activities of FtsA regulate the early establishment of a functional divisome during the Escherichia coli cell cycle
title_sort nucleotide-dependent activities of ftsa regulate the early establishment of a functional divisome during the escherichia coli cell cycle
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10213515/
https://www.ncbi.nlm.nih.gov/pubmed/37250038
http://dx.doi.org/10.3389/fmicb.2023.1171376
work_keys_str_mv AT morrisonjosiahj nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT ferreiracolbyn nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT silerevelynm nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT nelsonkatie nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT trebinocatherinee nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT pirainobenjamin nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle
AT cambergjodil nucleotidedependentactivitiesofftsaregulatetheearlyestablishmentofafunctionaldivisomeduringtheescherichiacolicellcycle