---
OA_place: publisher
_id: '20741'
abstract:
- lang: eng
text: "Life on Earth emerged when biomacromolecules were membrane-enclosed in a
confined space where many essential chemical reactions were more likely to happen
and thereby accelerate evolution. These kinds of membranes separated internal
reactions from the outside chaos while staying flexible so that those primordial
cells can move, adopt their shape and, most importantly, propagate. Such membrane
plasticity still remains a defining feature of all modern cell types. This remarkable
ability to change their shape is most prominently observed during their propagation
(i.e., cell division). Throughout division, a cell undergoes drastic change in
its shape, usually at the middle of the cell, pulling the two opposite membrane
sides inward, closer to each other, and, finally, culminating in pinching off
to separate the cell into two daughter cells. To achieve this, a cell needs to
employ a protein machinery, usually termed divisome, that can coordinate all necessary
intracellular processes with membrane remodelling and synthesis of other extracellular
structures that decorate a cell. The focus of this dissertation is a membrane-remodelling
FtsZ system that is present across all domains of life. FtsZ forms filaments that
further self-organize into ring-like structures at the cell septum and together
with other division proteins perform cell envelope synthesis and constriction.
However, there are still knowledge gaps in our mechanistic understanding of division
in both archaea and bacteria. My work presented in this dissertation centres around
a simple yet not well understood question: How is the divisome positioned correctly
at the mid-cell? To achieve the proper positioning, the divisome needs to (i)
be recruited to the mid-cell and (ii) localized orthogonally to the long cell
axis. I tackle these processes in two different systems by applying an in vitro
biochemical bottom-up reconstitution approach. I use purified components of Haloferax
volcanii and Escherichia coli divisome to explore how divisome is recruited to
the mid-cell in archaea and how the Z-ring positions orthogonally to the long
cell axis in bacteria, respectively. \r\n\r\nFirstly, I collaborate with archaeal
cell and structural biologists to explore the assembly of early division proteins
in two FtsZ-containing archaeon H. volcanii, a standard model system for understudied
archaeal organisms. I particularly address the hierarchy of interactions that
allow a tripartite complex formation (SepF-CdpB1-CdpB2) and how the hierarchy
of interactions ultimately leads to the recruitment of FtsZ filaments to the septum.
This part of work has been published in (Nußbaum et al., 2024). In collaboration
with evolutionary biologists, I shed light on ancient features that archaeal divisome
has retained to this day and also speculate on a property that it might have lost
during the course of evolution. \r\n\r\nNext, I switch my attention to E. coli
divisome. Particularly, I address the FtsZ’s intrinsic biophysical property that
drives the Z-ring diameter, and thereby the perpendicular orientation of the Z-ring
to the long cell axis based on suggested membrane curvature sensing mechanism
(Vanhille-Campos et al., 2024). This property allows formation of different Z-ring
diameters that match the variety of cell diameters present in prokaryotes. The
results showcase that the distribution of charged amino acids in the intrinsically
disordered linker at the C-terminus (CTL) of FtsZ is the major determining factor
of Z-ring diameter with inter-CTL interactions as an underlying mechanism. \r\n\r\nFinally,
I thoroughly explain the methodology I used to address the abovementioned projects,
and I finish with a discussion on how early archaeal divisome assembly and curvature
sensing mechanism in bacteria, at first sight unrelated topics, are interconnected
and important groundwork for both fundamental and translational research. "
acknowledged_ssus:
- _id: Bio
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Marko
full_name: Kojic, Marko
id: 73e7ecd4-dc85-11ea-9058-88a16394b160
last_name: Kojic
orcid: 0000-0001-7244-8128
citation:
ama: Kojic M. Towards understanding the assembly mechanisms of the Z-ring in Archaea
and Bacteria. 2025. doi:10.15479/AT-ISTA-20741
apa: Kojic, M. (2025). Towards understanding the assembly mechanisms of the Z-ring
in Archaea and Bacteria. Institute of Science and Technology Austria. https://doi.org/10.15479/AT-ISTA-20741
chicago: Kojic, Marko. “Towards Understanding the Assembly Mechanisms of the Z-Ring
in Archaea and Bacteria.” Institute of Science and Technology Austria, 2025. https://doi.org/10.15479/AT-ISTA-20741.
ieee: M. Kojic, “Towards understanding the assembly mechanisms of the Z-ring in
Archaea and Bacteria,” Institute of Science and Technology Austria, 2025.
ista: Kojic M. 2025. Towards understanding the assembly mechanisms of the Z-ring
in Archaea and Bacteria. Institute of Science and Technology Austria.
mla: Kojic, Marko. Towards Understanding the Assembly Mechanisms of the Z-Ring
in Archaea and Bacteria. Institute of Science and Technology Austria, 2025,
doi:10.15479/AT-ISTA-20741.
short: M. Kojic, Towards Understanding the Assembly Mechanisms of the Z-Ring in
Archaea and Bacteria, Institute of Science and Technology Austria, 2025.
corr_author: '1'
date_created: 2025-12-09T13:08:11Z
date_published: 2025-12-09T00:00:00Z
date_updated: 2026-04-07T12:27:58Z
day: '09'
ddc:
- '572'
degree_awarded: PhD
department:
- _id: GradSch
- _id: MaLo
doi: 10.15479/AT-ISTA-20741
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checksum: a3643d07e93134b2490a566b02a4517d
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date_created: 2025-12-10T13:09:58Z
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file_date_updated: 2025-12-10T13:09:58Z
has_accepted_license: '1'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '12'
oa: 1
oa_version: Published Version
publication_identifier:
isbn:
- 978-3-99078-073-2
issn:
- 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
record:
- id: '15118'
relation: part_of_dissertation
status: public
status: public
supervisor:
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
title: Towards understanding the assembly mechanisms of the Z-ring in Archaea and
Bacteria
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2025'
...
---
OA_place: publisher
_id: '14280'
abstract:
- lang: eng
text: "Cell division in Escherichia coli is performed by the divisome, a multi-protein
complex composed of more than 30 proteins. The divisome spans from the cytoplasm
through the inner membrane to the cell wall and the outer membrane. Divisome assembly
is initiated by a cytoskeletal structure, the so-called Z-ring, which localizes
at the center of the E. coli cell and determines the position of the future cell
septum. The Z-ring is composed of the highly conserved bacterial tubulin homologue
FtsZ, which forms treadmilling filaments. These filaments are recruited to the
inner membrane by FtsA, a highly conserved bacterial actin homologue. FtsA interacts
with other proteins in the periplasm and thus connects the cytoplasmic and periplasmic
components of the divisome. \r\nA previous model postulated that FtsA regulates
maturation of the divisome by switching from an oligomeric, inactive state to
a monomeric and active state. This model was based mostly on in vivo studies,
as a biochemical characterization of FtsA has been hampered by difficulties in
purifying the protein. Here, we studied FtsA using an in vitro reconstitution
approach and aimed to answer two questions: (i) How are dynamics from cytoplasmic,
treadmilling FtsZ filaments coupled to proteins acting in the periplasmic space
and (ii) How does FtsA regulate the maturation of the divisome?\r\nWe found that
the cytoplasmic peptides of the transmembrane proteins FtsN and FtsQ interact
directly with FtsA and can follow the spatiotemporal signal of FtsA/Z filaments.
When we investigated the underlying mechanism by imaging single molecules of FtsNcyto,
we found the peptide to interact transiently with FtsA. An in depth analysis of
the single molecule trajectories helped to postulate a model where PG synthases
follow the dynamics of FtsZ by a diffusion and capture mechanism. \r\nFollowing
up on these findings we were interested in how the self-interaction of FtsA changes
when it encounters FtsNcyto and if we can confirm the proposed oligomer-monomer
switch. For this, we compared the behavior of the previously identified, hyperactive
mutant FtsA R286W with wildtype FtsA. The mutant outperforms WT in mirroring and
transmitting the spatiotemporal signal of treadmilling FtsZ filaments. Surprisingly
however, we found that this was not due to a difference in the self-interaction
strength of the two variants, but a difference in their membrane residence time.
Furthermore, in contrast to our expectations, upon binding of FtsNcyto the measured
self-interaction of FtsA actually increased. \r\nWe propose that FtsNcyto induces
a rearrangement of the oligomeric architecture of FtsA. In further consequence
this change leads to more persistent FtsZ filaments which results in a defined
signalling zone, allowing formation of the mature divisome. The observed difference
between FtsA WT and R286W is due to the vastly different membrane turnover of
the proteins. R286W cycles 5-10x faster compared to WT which allows to sample
FtsZ filaments at faster frequencies. These findings can explain the observed
differences in toxicity for overexpression of FtsA WT and R286W and help to understand
how FtsA regulates divisome maturation."
acknowledged_ssus:
- _id: Bio
- _id: LifeSc
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Philipp
full_name: Radler, Philipp
id: 40136C2A-F248-11E8-B48F-1D18A9856A87
last_name: Radler
orcid: '0000-0001-9198-2182 '
citation:
ama: Radler P. Spatiotemporal signaling during assembly of the bacterial divisome.
2023. doi:10.15479/at:ista:14280
apa: Radler, P. (2023). Spatiotemporal signaling during assembly of the bacterial
divisome. Institute of Science and Technology Austria. https://doi.org/10.15479/at:ista:14280
chicago: Radler, Philipp. “Spatiotemporal Signaling during Assembly of the Bacterial
Divisome.” Institute of Science and Technology Austria, 2023. https://doi.org/10.15479/at:ista:14280.
ieee: P. Radler, “Spatiotemporal signaling during assembly of the bacterial divisome,”
Institute of Science and Technology Austria, 2023.
ista: Radler P. 2023. Spatiotemporal signaling during assembly of the bacterial
divisome. Institute of Science and Technology Austria.
mla: Radler, Philipp. Spatiotemporal Signaling during Assembly of the Bacterial
Divisome. Institute of Science and Technology Austria, 2023, doi:10.15479/at:ista:14280.
short: P. Radler, Spatiotemporal Signaling during Assembly of the Bacterial Divisome,
Institute of Science and Technology Austria, 2023.
corr_author: '1'
date_created: 2023-09-06T10:58:25Z
date_published: 2023-09-25T00:00:00Z
date_updated: 2026-04-07T14:06:05Z
day: '25'
ddc:
- '572'
degree_awarded: PhD
department:
- _id: GradSch
- _id: MaLo
doi: 10.15479/at:ista:14280
ec_funded: 1
file:
- access_level: closed
checksum: 87eef11fbc5c7df0826f12a3a629b444
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creator: pradler
date_created: 2023-10-04T10:11:53Z
date_updated: 2024-10-05T22:30:03Z
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file_id: '14390'
file_name: PhD Thesis_Philipp Radler_20231004.docx
file_size: 114932847
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content_type: application/pdf
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date_created: 2023-10-04T10:11:21Z
date_updated: 2024-10-05T22:30:03Z
embargo: 2024-10-04
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file_size: 37838778
relation: main_file
file_date_updated: 2024-10-05T22:30:03Z
has_accepted_license: '1'
keyword:
- Cell Division
- Reconstitution
- FtsZ
- FtsA
- Divisome
- E.coli
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
page: '156'
project:
- _id: 2595697A-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '679239'
name: Self-Organization of the Bacterial Cell
- _id: fc38323b-9c52-11eb-aca3-ff8afb4a011d
grant_number: P34607
name: In vitro reconstitution of bacterial cell division
- _id: 2596EAB6-B435-11E9-9278-68D0E5697425
grant_number: ALTF 2015-1163
name: Synthesis of bacterial cell wall
- _id: 259B655A-B435-11E9-9278-68D0E5697425
grant_number: LT000824/2016
name: Reconstitution of bacterial cell wall synthesis
publication_identifier:
isbn:
- 978-3-99078-033-6
issn:
- 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
record:
- id: '10934'
relation: research_data
status: public
- id: '11373'
relation: part_of_dissertation
status: public
- id: '7387'
relation: part_of_dissertation
status: public
status: public
supervisor:
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
title: Spatiotemporal signaling during assembly of the bacterial divisome
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2023'
...
---
OA_place: publisher
_id: '14510'
abstract:
- lang: eng
text: "Clathrin-mediated endocytosis (CME) is vital for the regulation of plant
growth and\r\ndevelopment by controlling plasma membrane protein composition and
cargo uptake. CME\r\nrelies on the precise recruitment control of protein regulators
for vesicle maturation and\r\nrelease. During the early stages of endocytosis,
an area of flat membrane is remodelled by\r\nproteins to create a spherical vesicle
against intracellular forces. After the Clathrin-coated\r\nvesicle (CCV) is fully
formed, scission machinery releases it from the plasma membrane,\r\nand cargo
proceeds for recycling or degradation through early endosomes / Trans Golgi\r\nnetwork.
Protein machineries that mediate membrane bending and vesicle release in plants\r\nare
unknown. However, studies show, that plant endocytosis is actin independent, thus\r\nindicating
that plants utilize a unique mechanism to mediate membrane bending against highturgor
pressure compared to other model systems. First, by using biochemical and advanced\r\nlive
microscopy approaches we investigate the TPLATE complex, a plant-specific\r\nendocytosis
protein complex. We found that TPLATE is peripherally associated with\r\nclathrin-coated
vesicles and localises at the rim of endocytosis events. Next, our study of\r\nplant
Dynamin-related protein 1C (DRP1C), which was hypothesised previously to play
a\r\nrole in vesicle release, shows the recruitment of the protein already at
the early stages of\r\nendocytosis. Moreover, DRP1C assembles into organised ring-like
structures and is able to\r\ninduce membrane deformation and tubulation, suggesting
its role also in membrane bending\r\nduring early CME. Based on the data from
mammalian and yeast systems, plant DynaminRelated Proteins 2 and SH3P2 protein
are strong candidates to be part of the plant vesicle\r\nscission machinery; however,
their precise role in plant CME has not been yet elucidated.\r\nHere, we characterised
DRP2s and SH3P2 roles in CME by combining high-resolution\r\nimaging of endocytic
events in vivo and protein characterisation. Although DRP2s and\r\nSH3P2 arrive
together during late CME and physically interact, genetic analysis using\r\n∆sh3p1,2,3
mutant and complementation with non-DRP2-interacting SH3P2 variants suggest\r\nthat
SH3P2 does not directly recruit DRP2s to the site of endocytosis. Summarising
our\r\nresearch, these observations provide new important insights into the mechanism
of plant\r\nCME and show that, despite plants posses many homologues of mammalian
and yeast CME\r\ncomponents, they do not necessarily act in the same manner. "
acknowledged_ssus:
- _id: EM-Fac
- _id: Bio
- _id: LifeSc
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Nataliia
full_name: Gnyliukh, Nataliia
id: 390C1120-F248-11E8-B48F-1D18A9856A87
last_name: Gnyliukh
orcid: 0000-0002-2198-0509
citation:
ama: Gnyliukh N. Mechanism of clathrin-coated vesicle formation during endocytosis
in plants. 2023. doi:10.15479/at:ista:14510
apa: Gnyliukh, N. (2023). Mechanism of clathrin-coated vesicle formation during
endocytosis in plants. Institute of Science and Technology Austria. https://doi.org/10.15479/at:ista:14510
chicago: Gnyliukh, Nataliia. “Mechanism of Clathrin-Coated Vesicle Formation during
Endocytosis in Plants.” Institute of Science and Technology Austria, 2023. https://doi.org/10.15479/at:ista:14510.
ieee: N. Gnyliukh, “Mechanism of clathrin-coated vesicle formation during endocytosis
in plants,” Institute of Science and Technology Austria, 2023.
ista: Gnyliukh N. 2023. Mechanism of clathrin-coated vesicle formation during endocytosis
in plants. Institute of Science and Technology Austria.
mla: Gnyliukh, Nataliia. Mechanism of Clathrin-Coated Vesicle Formation during
Endocytosis in Plants. Institute of Science and Technology Austria, 2023,
doi:10.15479/at:ista:14510.
short: N. Gnyliukh, Mechanism of Clathrin-Coated Vesicle Formation during Endocytosis
in Plants, Institute of Science and Technology Austria, 2023.
corr_author: '1'
date_created: 2023-11-10T09:10:06Z
date_published: 2023-11-10T00:00:00Z
date_updated: 2026-05-04T22:30:33Z
day: '10'
ddc:
- '570'
degree_awarded: PhD
department:
- _id: GradSch
- _id: JiFr
- _id: MaLo
doi: 10.15479/at:ista:14510
ec_funded: 1
file:
- access_level: closed
checksum: 3d5e680bfc61f98e308c434f45cc9bd6
content_type: application/vnd.openxmlformats-officedocument.wordprocessingml.document
creator: ngnyliuk
date_created: 2023-11-20T09:18:51Z
date_updated: 2024-11-23T23:30:38Z
embargo_to: open_access
file_id: '14567'
file_name: Thesis_Gnyliukh_final_08_11_23.docx
file_size: 20824903
relation: source_file
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checksum: bfc96d47fc4e7e857dd71656097214a4
content_type: application/pdf
creator: ngnyliuk
date_created: 2023-11-20T09:23:11Z
date_updated: 2024-11-23T23:30:38Z
embargo: 2024-11-23
file_id: '14568'
file_name: Thesis_Gnyliukh_final_20_11_23.pdf
file_size: 24871844
relation: main_file
file_date_updated: 2024-11-23T23:30:38Z
has_accepted_license: '1'
keyword:
- Clathrin-Mediated Endocytosis
- vesicle scission
- Dynamin-Related Protein 2
- SH3P2
- TPLATE complex
- Total internal reflection fluorescence microscopy
- Arabidopsis thaliana
language:
- iso: eng
month: '11'
oa: 1
oa_version: Published Version
page: '180'
project:
- _id: 2564DBCA-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '665385'
name: International IST Doctoral Program
publication_identifier:
isbn:
- 978-3-99078-037-4
issn:
- 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
record:
- id: '14591'
relation: part_of_dissertation
status: public
- id: '9887'
relation: part_of_dissertation
status: public
- id: '8139'
relation: part_of_dissertation
status: public
status: public
supervisor:
- first_name: Jiří
full_name: Friml, Jiří
id: 4159519E-F248-11E8-B48F-1D18A9856A87
last_name: Friml
orcid: 0000-0002-8302-7596
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
title: Mechanism of clathrin-coated vesicle formation during endocytosis in plants
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2023'
...
---
OA_place: publisher
_id: '8341'
abstract:
- lang: eng
text: "One of the most striking hallmarks of the eukaryotic cell is the presence
of intracellular vesicles and organelles. Each of these membrane-enclosed compartments
has a distinct composition of lipids and proteins, which is essential for accurate
membrane traffic and homeostasis. Interestingly, their biochemical identities
are achieved with the help\r\nof small GTPases of the Rab family, which cycle
between GDP- and GTP-bound forms on the selected membrane surface. While this
activity switch is well understood for an individual protein, how Rab GTPases
collectively transition between states to generate decisive signal propagation
in space and time is unclear. In my PhD thesis, I present\r\nin vitro reconstitution
experiments with theoretical modeling to systematically study a minimal Rab5 activation
network from bottom-up. We find that positive feedback based on known molecular
interactions gives rise to bistable GTPase activity switching on system’s scale.
Furthermore, we determine that collective transition near the critical\r\npoint
is intrinsically stochastic and provide evidence that the inactive Rab5 abundance
on the membrane can shape the network response. Finally, we demonstrate that collective
switching can spread on the lipid bilayer as a traveling activation wave, representing
a possible emergent activity pattern in endosomal maturation. Together, our\r\nfindings
reveal new insights into the self-organization properties of signaling networks
away from chemical equilibrium. Our work highlights the importance of systematic
characterization of biochemical systems in well-defined physiological conditions.
This way, we were able to answer long-standing open questions in the field and
close the gap between regulatory processes on a molecular scale and emergent responses
on system’s level."
acknowledged_ssus:
- _id: Bio
- _id: LifeSc
- _id: NanoFab
acknowledgement: My thanks goes to the Loose lab members, BioImaging, Life Science
and Nanofabrication Facilities and the wonderful international community at IST
for sharing this experience with me.
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Urban
full_name: Bezeljak, Urban
id: 2A58201A-F248-11E8-B48F-1D18A9856A87
last_name: Bezeljak
orcid: 0000-0003-1365-5631
citation:
ama: Bezeljak U. In vitro reconstitution of a Rab activation switch. 2020. doi:10.15479/AT:ISTA:8341
apa: Bezeljak, U. (2020). In vitro reconstitution of a Rab activation switch.
Institute of Science and Technology Austria. https://doi.org/10.15479/AT:ISTA:8341
chicago: Bezeljak, Urban. “In Vitro Reconstitution of a Rab Activation Switch.”
Institute of Science and Technology Austria, 2020. https://doi.org/10.15479/AT:ISTA:8341.
ieee: U. Bezeljak, “In vitro reconstitution of a Rab activation switch,” Institute
of Science and Technology Austria, 2020.
ista: Bezeljak U. 2020. In vitro reconstitution of a Rab activation switch. Institute
of Science and Technology Austria.
mla: Bezeljak, Urban. In Vitro Reconstitution of a Rab Activation Switch.
Institute of Science and Technology Austria, 2020, doi:10.15479/AT:ISTA:8341.
short: U. Bezeljak, In Vitro Reconstitution of a Rab Activation Switch, Institute
of Science and Technology Austria, 2020.
corr_author: '1'
date_created: 2020-09-08T08:53:53Z
date_published: 2020-09-08T00:00:00Z
date_updated: 2026-04-08T07:24:56Z
day: '08'
ddc:
- '570'
degree_awarded: PhD
department:
- _id: MaLo
doi: 10.15479/AT:ISTA:8341
file:
- access_level: closed
checksum: 70871b335a595252a66c6bbf0824fb02
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creator: dernst
date_created: 2020-09-08T09:00:29Z
date_updated: 2021-09-16T12:49:12Z
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creator: dernst
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month: '09'
oa: 1
oa_version: Published Version
page: '215'
publication_identifier:
issn:
- 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
record:
- id: '7580'
relation: part_of_dissertation
status: public
status: public
supervisor:
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
title: In vitro reconstitution of a Rab activation switch
tmp:
image: /images/cc_by_nc_sa.png
legal_code_url: https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC
BY-NC-SA 4.0)
short: CC BY-NC-SA (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2020'
...
---
OA_place: publisher
_id: '8358'
abstract:
- lang: eng
text: "During bacterial cell division, the tubulin-homolog FtsZ forms a ring-like
structure at the center of the cell. This so-called Z-ring acts as a scaffold
recruiting several division-related proteins to mid-cell and plays a key role
in distributing proteins at the division site, a feature driven by the treadmilling
motion of FtsZ filaments around the septum. What regulates the architecture, dynamics
and stability of the Z-ring is still poorly understood, but FtsZ-associated proteins
(Zaps) are known to play an important role. \r\nAdvances in fluorescence microscopy
and in vitro reconstitution experiments have helped to shed light into some of
the dynamic properties of these complex systems, but methods that allow to collect
and analyze large quantitative data sets of the underlying polymer dynamics are
still missing.\r\nHere, using an in vitro reconstitution approach, we studied
how different Zaps affect FtsZ filament dynamics and organization into large-scale
patterns, giving special emphasis to the role of the well-conserved protein ZapA.
For this purpose, we use high-resolution fluorescence microscopy combined with
novel image analysis workfows to study pattern organization and polymerization
dynamics of active filaments. We quantified the influence of Zaps on FtsZ on three
diferent spatial scales: the large-scale organization of the membrane-bound filament
network, the underlying\r\npolymerization dynamics and the behavior of single
molecules.\r\nWe found that ZapA cooperatively increases the spatial order of
the filament network, binds only transiently to FtsZ filaments and has no effect
on filament length and treadmilling velocity. Our data provides a model for how
FtsZ-associated proteins can increase the precision and stability of the bacterial
cell division machinery in a\r\nswitch-like manner, without compromising filament
dynamics. Furthermore, we believe that our automated quantitative methods can
be used to analyze a large variety of dynamic cytoskeletal systems, using standard
time-lapse\r\nmovies of homogeneously labeled proteins obtained from experiments
in vitro or even inside the living cell.\r\n"
acknowledged_ssus:
- _id: Bio
acknowledgement: I should also express my gratitude to the bioimaging facility at
IST Austria, for their assistance with the TIRF setup over the years, and especially
to Christoph Sommer, who gave me a lot of input when I was starting to dive into
programming.
alternative_title:
- ISTA Thesis
article_processing_charge: No
author:
- first_name: Paulo R
full_name: Dos Santos Caldas, Paulo R
id: 38FCDB4C-F248-11E8-B48F-1D18A9856A87
last_name: Dos Santos Caldas
orcid: 0000-0001-6730-4461
citation:
ama: Dos Santos Caldas PR. Organization and dynamics of treadmilling filaments in
cytoskeletal networks of FtsZ and its crosslinkers. 2020. doi:10.15479/AT:ISTA:8358
apa: Dos Santos Caldas, P. R. (2020). Organization and dynamics of treadmilling
filaments in cytoskeletal networks of FtsZ and its crosslinkers. Institute
of Science and Technology Austria. https://doi.org/10.15479/AT:ISTA:8358
chicago: Dos Santos Caldas, Paulo R. “Organization and Dynamics of Treadmilling
Filaments in Cytoskeletal Networks of FtsZ and Its Crosslinkers.” Institute of
Science and Technology Austria, 2020. https://doi.org/10.15479/AT:ISTA:8358.
ieee: P. R. Dos Santos Caldas, “Organization and dynamics of treadmilling filaments
in cytoskeletal networks of FtsZ and its crosslinkers,” Institute of Science and
Technology Austria, 2020.
ista: Dos Santos Caldas PR. 2020. Organization and dynamics of treadmilling filaments
in cytoskeletal networks of FtsZ and its crosslinkers. Institute of Science and
Technology Austria.
mla: Dos Santos Caldas, Paulo R. Organization and Dynamics of Treadmilling Filaments
in Cytoskeletal Networks of FtsZ and Its Crosslinkers. Institute of Science
and Technology Austria, 2020, doi:10.15479/AT:ISTA:8358.
short: P.R. Dos Santos Caldas, Organization and Dynamics of Treadmilling Filaments
in Cytoskeletal Networks of FtsZ and Its Crosslinkers, Institute of Science and
Technology Austria, 2020.
corr_author: '1'
date_created: 2020-09-10T09:26:49Z
date_published: 2020-09-10T00:00:00Z
date_updated: 2026-04-08T07:26:30Z
day: '10'
ddc:
- '572'
degree_awarded: PhD
department:
- _id: MaLo
doi: 10.15479/AT:ISTA:8358
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has_accepted_license: '1'
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
page: '135'
publication_identifier:
isbn:
- 978-3-99078-009-1
issn:
- 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
record:
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relation: part_of_dissertation
status: public
- id: '7572'
relation: dissertation_contains
status: public
status: public
supervisor:
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
title: Organization and dynamics of treadmilling filaments in cytoskeletal networks
of FtsZ and its crosslinkers
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2020'
...