Direct Arp2/3-vinculin binding is essential for cell spreading, but only on compliant substrates and in 3D
Posted on: 15 September 2019 , updated on: 24 September 2019
Preprint posted on 4 September 2019
Categories: cancer biology, cell biology
Context:
The actin cytoskeleton is actively remodeled in response to differential extra-cellular matrix (ECM) cues. Thus, the architectural and the chemical composition of ECM can regulate complex cell shapes. How cells respond to the different ECM environment are one of the major questions in the field. It is also becoming clear that response of cells to mechanochemical changes encountered in2D and 3D environments occurs through adhesion complexes with different mechanisms in place (Doyle and Yamada et al 2015).
Major findings:
To understand how cells respond to different ECM environments, i.e. on stiff 2D planar and 3D reconstituted soft environments, the authors performed a CRISPR-Cas9 knock-out screen of actin binding proteins (ABPs) and then characterized the cell shapes after cells were plated on the planar surfaces like glass (2D) and also in 3D collagen matrices. Knock-out of actin nucleators FMNL1 and Arp3 revealed difference in the spreading between 2D and 3D. In order to understand if the architecture of the fibrillar collagen or the stiffness of the ECM is important for the spreading of the cells, authors looked at the spreading of cells on the collagen coated on the glass and also silicone-based substrates with similar stiffness like that of collagen but without any fibrils. In both cases Arp3 knock out showed reduced spreading, which suggested that spreading is a combination of stiffness and architectural features. In 3D, cells show elongated adhesion compared to Arp3 KO cells, whereas in 2D no such differences were seen. Authors suggest the presence of a compensatory pathways in 2D, and also show that a phospho-specific Arp3-vinculin interaction is important for cells to spread in soft, 3D collagen environments.
What I like about the pre-print:
This work along with others shed light on the how cells sense their micro-environment. Cells on 2D form more lamellopodial like protrusions compared to 3D. It is also very clear from the experiments that cells respond to the difference in the stiffness of the substrate, which is uniform in 2D and also to the architectural make of the 3D like the fibers of collagen. It is becoming increasingly clear that the cells mechano-sensing on the 2D substrates involves dynamic interactions between vinculin, talin and actin filaments generating a molecular clutch (Hu et al., 2007). It is not clear if there are difference in the mechano-sensing mechanisms between 2D and 3D, and we have only begun to understand the differences that might exist.
What’s next and my questions to the authors:
Cells in 2D largely depend on the ECM stiffness and those in 3D depend on the fibril architecture, cross-linking, ECM adhesion ligand density and pore size of the fibrils. This makes it difficult to compare between 2D and 3D ECM. There lies a large conundrum about the 3D matrix where the overall macro gel stiffness differs from the fiber stiffness that cells witness in 3D (Guthold et al. 2007). I wonder how authors plan to answer this, if the difference in the spreading difference they see between 2D and 3D is because of the fibril stiffness that is seen by the cells? Vinculin knock-out in MEFs have been shown to change morphology in 3D but not in 2D (Thievessen et al., 2015). It will be interesting to see if the binding partners of vinculin changes based on the conformation it shows in 2D versus 3D. This could be a result of the Rho/Rac activity in the cells (Deakin et al 2011). Vinculin mutant where it does not interact with Arp3 show diminished lamellipodia and spreading on fibronectin in 2D (DeMali et al, 2002) similar to what authors have in 3D. It would be interesting to see if this could be a combination of the available adhesive ligands in 2D and stiffness together effects the spreading of cells. Authors mention at the different mechanistic pathways and mechanisms in 2D and 3D, it would be great if any of the of the components of these pathways were tested in future. For example, does the clutch model exist in 3D?
doi: https://doi.org/10.1242/prelights.13860
Read preprint
(No Ratings Yet)Sign up to customise the site to your preferences and to receive alerts
Register hereAlso in the cancer biology category:
Integrin conformation-dependent neutrophil slowing obstructs the capillaries of the pre-metastatic lung in a model of breast cancer
Simon Cleary
Mitochondria-derived nuclear ATP surge protects against confinement-induced proliferation defects
Teodora Piskova
Spatial transcriptomics elucidates medulla niche supporting germinal center response in myasthenia gravis thymoma
Jessica Chevallier
Also in the cell biology category:
Motor Clustering Enhances Kinesin-driven Vesicle Transport
Sharvari Pitke
Cellular signalling protrusions enable dynamic distant contacts in spinal cord neurogenesis
Ankita Walvekar
Green synthesized silver nanoparticles from Moringa: Potential for preventative treatment of SARS-CoV-2 contaminated water
Safieh Shah, Benjamin Dominik Maier
preLists in the cancer biology category:
BSCB-Biochemical Society 2024 Cell Migration meeting
This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.
| List by | Reinier Prosee |
CSHL 87th Symposium: Stem Cells
Preprints mentioned by speakers at the #CSHLsymp23
| List by | Alex Eve |
Journal of Cell Science meeting ‘Imaging Cell Dynamics’
This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.
| List by | Helen Zenner |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
| List by | Nadja Hümpfer et al. |
Fibroblasts
The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!
| List by | Osvaldo Contreras |
Single Cell Biology 2020
A list of preprints mentioned at the Wellcome Genome Campus Single Cell Biology 2020 meeting.
| List by | Alex Eve |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
| List by | Madhuja Samaddar et al. |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
| List by | Rob Hynds |
Anticancer agents: Discovery and clinical use
Preprints that describe the discovery of anticancer agents and their clinical use. Includes both small molecules and macromolecules like biologics.
| List by | Zhang-He Goh |
Biophysical Society Annual Meeting 2019
Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA
| List by | Joseph Jose Thottacherry |
Also in the cell biology category:
BSCB-Biochemical Society 2024 Cell Migration meeting
This preList features preprints that were discussed and presented during the BSCB-Biochemical Society 2024 Cell Migration meeting in Birmingham, UK in April 2024. Kindly put together by Sara Morais da Silva, Reviews Editor at Journal of Cell Science.
| List by | Reinier Prosee |
‘In preprints’ from Development 2022-2023
A list of the preprints featured in Development's 'In preprints' articles between 2022-2023
| List by | Alex Eve, Katherine Brown |
preLights peer support – preprints of interest
This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.
| List by | preLights peer support |
The Society for Developmental Biology 82nd Annual Meeting
This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.
| List by | Joyce Yu, Katherine Brown |
CSHL 87th Symposium: Stem Cells
Preprints mentioned by speakers at the #CSHLsymp23
| List by | Alex Eve |
Journal of Cell Science meeting ‘Imaging Cell Dynamics’
This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.
| List by | Helen Zenner |
9th International Symposium on the Biology of Vertebrate Sex Determination
This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.
| List by | Martin Estermann |
Alumni picks – preLights 5th Birthday
This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.
| List by | Sergio Menchero et al. |
CellBio 2022 – An ASCB/EMBO Meeting
This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.
| List by | Nadja Hümpfer et al. |
Fibroblasts
The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!
| List by | Osvaldo Contreras |
EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)
A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.
| List by | Alex Eve |
FENS 2020
A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020
| List by | Ana Dorrego-Rivas |
Planar Cell Polarity – PCP
This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.
| List by | Ana Dorrego-Rivas |
BioMalPar XVI: Biology and Pathology of the Malaria Parasite
[under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria
| List by | Dey Lab, Samantha Seah |
1
Cell Polarity
Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.
| List by | Yamini Ravichandran |
TAGC 2020
Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20
| List by | Maiko Kitaoka et al. |
3D Gastruloids
A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.
| List by | Paul Gerald L. Sanchez and Stefano Vianello |
ECFG15 – Fungal biology
Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome
| List by | Hiral Shah |
ASCB EMBO Annual Meeting 2019
A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)
| List by | Madhuja Samaddar et al. |
EMBL Seeing is Believing – Imaging the Molecular Processes of Life
Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019
| List by | Dey Lab |
Autophagy
Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.
| List by | Sandra Malmgren Hill |
Lung Disease and Regeneration
This preprint list compiles highlights from the field of lung biology.
| List by | Rob Hynds |
Cellular metabolism
A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.
| List by | Pablo Ranea Robles |
BSCB/BSDB Annual Meeting 2019
Preprints presented at the BSCB/BSDB Annual Meeting 2019
| List by | Dey Lab |
MitoList
This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.
| List by | Sandra Franco Iborra |
ASCB/EMBO Annual Meeting 2018
This list relates to preprints that were discussed at the recent ASCB conference.
| List by | Dey Lab, Amanda Haage |