Molecular motion and tridimensional nanoscale localization of kindlin control integrin activation in focal adhesions

Thomas Orré, Zeynep Karatas, Birgit Kastberger, Clément Cabriel, Ralph T. Böttcher, Sandrine Lévêque-Fort, Jean-Baptiste Sibarita, Reinhard Fässler, Bernhard Wehrle-Haller, Olivier Rossier, Grégory Giannone

Posted on: 18 September 2020

Preprint posted on 6 August 2020

Article now published in Nature Communications at

Talins and Kindlins: partners in crime with specific modes of transportation.

Selected by Amanda Haage

Categories: cell biology

Why This Is Cool – This article is all about providing detailed localization data to the integrin adhesome. The “adhesome” refers to the many proteins that cluster around the cytoplasmic tails of integrins and mediate multiple signaling outcomes and links the complex to the actin cytoskeleton. In 2D cell culture this adhesome is pretty much used interchangeably with focal adhesions (FAs), and previous work has shown that there are distinct layers to FAs extending inward from the membrane (1). Here, the authors utilize cutting edge microscopy techniques to track single proteins, namely integrins, talins, and kindlins, in and out of FAs. By first tracking integrins, they show that they become immobilized in FAs and there are slight differences, where ß3 immobilization seems to be more dependent on Talin, while ß1 requires both Talin and Kindlin. These differences are the start of a line of evidence used to connect differences in diffusion/immobilization of Kindlin with a specific role in FAs. Tracking Kindlin in conjunction with various point mutations shows that unlike Talin (2), Kindlin is in free diffusion along the membrane driven by its unique PH domain. Kindlin binding to integrins mediates its immobilization in FAs and it remains near the membrane and integrin cytoplasmic tails instead of repositioning inward similar to vinculin (3). Ending with function, the PH domain and free diffusion along the membrane of Kindlin seems to be fundamental to promoting integrin activation and cell spreading.


Figure1: Membrane recruitment and diffusion of kindlin‐2 is crucial during cell spreading. Top: Phase contrast images showing the morphological features of cells classified as non‐spread, partially spread, or spread. Bottom: Relative fraction of non‐spread, partially spread, and spread KindKo cells 2 days after re‐expression of kindlin‐2‐WT or mutated variants and 4h after seeding on fibronectin. Values represent the average of fractions from 3 independent experiments (error bars: SEM). GFP‐ paxillin: 141 cells; GFP‐paxillin + mEos2‐kindlin‐2‐WT: 312 cells; GFP‐paxillin + mEos2‐kindlin‐2‐ΔPH: 311 cells; GFP‐paxillin + mEos2‐kindlin‐2‐ΔPH‐CAAX: 309 cells; GFP‐paxillin + mEos2‐kindlin‐2‐ QW615/615AA: 284 cells; GFP‐paxillin + mEos2‐kindlin‐2‐WT‐CAAX: 289 cells. Between 13 and 41 fields of view (20x objective) per condition and per experiment were used to quantify cell spreading, except for the cells expressing GFP‐paxillin alone (negative control: between 8 and 17 fields per experience). Between 84 and 107 cells per condition and per experiment were included (except for the GFP‐paxillin condition: between 31 and 58 cells per experiment).



Why I Selected It – One of the influential reviews in my field that I never forget the name of is “Talins and Kindlins: partners in integrin-mediated adhesion”(4), because in my head I have always read it as “Talins and Kindlins: partners in crime.” Essentially it reviews the well-established theory that both the FERM containing adaptors, Talin and Kindlin, bind to the cytoplasmic tails of integrins and facilitate activation, signaling, and linkage to the cytoskeleton. Talin-1’s role has been investigated more thoroughly (5), but the overlapping roles of these proteins remains an open question. What this study does really well is provide detailed and convincing data for distinct functions in FAs via distinct localizations and trafficking between the two.


Open Questions –

  1. For the functional experiments, Kindlin knockout cells are used to demonstrate rescue, but could the other results be influenced by the various expression levels induced with transfected constructs?
  2. Though the differing recruitment of Talin is cited, can you also replicate these important results with your system?
  3. Here you are focusing on the classical proteins of the field, i.e. ß3/ß1 integrins, Talin-1, Kindlin-2, but how much of this work is applicable to the increased complexity of the adhesome? Say Talin-2, the other Kindlins, and the other integrins.


Related References –

  1. Distinct Layers in FAs
    1. Kanchanawong, P. et al. Nanoscale architecture of integrin‐based cell adhesions. Nature 468, 580–584 (2010).
  1. Talin Recruitment
    1. Rossier, O. et al. Integrins β1 and β3 exhibit distinct dynamic nanoscale organizations inside focal adhesions. Cell Biol. 14, 1057–67 (2012).
  1. Vinculin Organization
    1. Case, L. B. et al. Molecular mechanism of vinculin activation and nanoscale spatial organization in focal adhesions. Cell Biol. 17, 880–892 (2015).
  1. Talins and Kindlins
    1. Calderwood, D. a, Campbell, I. D. & Critchley, D. R. Talins and kindlins: partners in integrin‐ mediated adhesion. Rev. Mol. Cell Biol. 14, 503–17 (2013).
  1. Talin the Master Regulator
    1. Klapholz, B. & Brown, N.H. Talin – the master of integrin adhesions. Cell Sci 130(15): 2435 – 2446.



Read preprint (No Ratings Yet)

Have your say

Your email address will not be published. Required fields are marked *

This site uses Akismet to reduce spam. Learn how your comment data is processed.

Sign up to customise the site to your preferences and to receive alerts

Register here

Also in the cell biology category:

Mitochondria-derived nuclear ATP surge protects against confinement-induced proliferation defects

Ritobrata Ghose, Fabio Pezzano, Savvas Kourtis, et al.

Selected by 16 May 2024

Teodora Piskova

Cell Biology

Lipid-Based Transfection of Zebrafish Embryos: A Robust Protocol for Nucleic Acid Delivery

Aslihan Terzi, Tiger Liao, Adrian Jacobo

Selected by 09 May 2024

Roberto Rodríguez-Morales


Fetal brain response to maternal inflammation requires microglia

Bridget Elaine LaMonica Ostrem, Nuria Dominguez Iturza, Jeffrey Stogsdill, et al.

Selected by 24 April 2024

Manuel Lessi


preLists 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.


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


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


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


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

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

ASCB/EMBO Annual Meeting 2018

This list relates to preprints that were discussed at the recent ASCB conference.


List by Dey Lab, Amanda Haage