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Essential and dispensable domains of DivIVA for walled growth in filamentous Actinomycetota

Lubbers, Maarten LU ; Bajramović, Belmin ; Ongenae, Véronique ; Willemse, Joost ; de Bruin, Dieuwertje ; Mulder, Niels ; Zhang, Le ; Vriesendorp, Bastienne ; Barona-Gomez, Francisco and Briegel, Ariane , et al. (2025) In Open biology 15(11).
Abstract

The morphogenetic protein DivIVA exhibits diverse functions across bacterial phyla. In Bacillota, DivIVA is primarily involved in cell division, whereas in Actinomycetota, it plays a central role in coordinating polar growth. Due to its essentiality in Actinomycetota, gaining insight into its structural functions is challenging. We studied truncated DivIVA proteins using a unique divIVA deletion mutant in cell wall-deficient Kitasatospora viridifaciens L-forms. DivIVA comprises an N-terminal domain consisting of a coiled-coil segment bearing a membrane-targeting structure at the N-terminal end, followed by an intercoil region, a larger coiled-coil and a C-terminal domain. Deleting either the intercoil or C-terminal region affected... (More)

The morphogenetic protein DivIVA exhibits diverse functions across bacterial phyla. In Bacillota, DivIVA is primarily involved in cell division, whereas in Actinomycetota, it plays a central role in coordinating polar growth. Due to its essentiality in Actinomycetota, gaining insight into its structural functions is challenging. We studied truncated DivIVA proteins using a unique divIVA deletion mutant in cell wall-deficient Kitasatospora viridifaciens L-forms. DivIVA comprises an N-terminal domain consisting of a coiled-coil segment bearing a membrane-targeting structure at the N-terminal end, followed by an intercoil region, a larger coiled-coil and a C-terminal domain. Deleting either the intercoil or C-terminal region affected branching. We also created a minimized variant in which both were deleted simultaneously, retaining the N-terminal domain and the second coiled-coil. Expression of this variant caused severe growth defects. Cells showed increased hyphal width, thicker cell walls and frequent tip bursting. Finally, we successfully introduced chimeric DivIVA from the unicellular actinobacterium Mycolicibacterium smegmatis with the membrane targeting domain of K. viridifaciens DivIVA, demonstrating functional conservation within the phylum. By contrast, chimeric DivIVA proteins from Bacillus subtilis could not support growth, underscoring that polar growth is encoded in Actinomycetota-specific amino acid motifs in the first and second coiled-coils. These findings enhance our understanding of the structure–function relationship for DivIVA and present new opportunities to study polar growth.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cell wall deficiency, hyphal growth, L-forms, morphogenesis, Streptomycetaceae
in
Open biology
volume
15
issue
11
article number
250213
publisher
Royal Society Publishing
external identifiers
  • scopus:105020894120
  • pmid:41187904
ISSN
2046-2441
DOI
10.1098/rsob.250213
language
English
LU publication?
yes
additional info
Publisher Copyright: © 2025 Royal Society Publishing. All rights reserved.
id
b9bb6f59-ca81-43b9-a269-2a87f55f81c8
date added to LUP
2025-12-11 13:36:17
date last changed
2025-12-12 03:00:11
@article{b9bb6f59-ca81-43b9-a269-2a87f55f81c8,
  abstract     = {{<p>The morphogenetic protein DivIVA exhibits diverse functions across bacterial phyla. In Bacillota, DivIVA is primarily involved in cell division, whereas in Actinomycetota, it plays a central role in coordinating polar growth. Due to its essentiality in Actinomycetota, gaining insight into its structural functions is challenging. We studied truncated DivIVA proteins using a unique divIVA deletion mutant in cell wall-deficient Kitasatospora viridifaciens L-forms. DivIVA comprises an N-terminal domain consisting of a coiled-coil segment bearing a membrane-targeting structure at the N-terminal end, followed by an intercoil region, a larger coiled-coil and a C-terminal domain. Deleting either the intercoil or C-terminal region affected branching. We also created a minimized variant in which both were deleted simultaneously, retaining the N-terminal domain and the second coiled-coil. Expression of this variant caused severe growth defects. Cells showed increased hyphal width, thicker cell walls and frequent tip bursting. Finally, we successfully introduced chimeric DivIVA from the unicellular actinobacterium Mycolicibacterium smegmatis with the membrane targeting domain of K. viridifaciens DivIVA, demonstrating functional conservation within the phylum. By contrast, chimeric DivIVA proteins from Bacillus subtilis could not support growth, underscoring that polar growth is encoded in Actinomycetota-specific amino acid motifs in the first and second coiled-coils. These findings enhance our understanding of the structure–function relationship for DivIVA and present new opportunities to study polar growth.</p>}},
  author       = {{Lubbers, Maarten and Bajramović, Belmin and Ongenae, Véronique and Willemse, Joost and de Bruin, Dieuwertje and Mulder, Niels and Zhang, Le and Vriesendorp, Bastienne and Barona-Gomez, Francisco and Briegel, Ariane and van Wezel, Gilles P. and Flärdh, Klas and Claessen, Dennis}},
  issn         = {{2046-2441}},
  keywords     = {{cell wall deficiency; hyphal growth; L-forms; morphogenesis; Streptomycetaceae}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{11}},
  publisher    = {{Royal Society Publishing}},
  series       = {{Open biology}},
  title        = {{Essential and dispensable domains of DivIVA for walled growth in filamentous Actinomycetota}},
  url          = {{http://dx.doi.org/10.1098/rsob.250213}},
  doi          = {{10.1098/rsob.250213}},
  volume       = {{15}},
  year         = {{2025}},
}