Genetically encoded reporters of actin filament organization in living cells and tissues

Carla Silva Martins (1) , Francois Iv (1) , Shashi Kumar Suman (2) , Thomas Panagiotou (3) , Clara Sidor (4) , Maria Ruso-Lopez (5) , Camille Plancke (2) , Shizue Omi (1) , Maxime Gomes (1) , Alexander Llewellyn (1) , Sourish Reddy Bandi (1) , Laurie Ramond (1) , Federica Arbizzani (6) , Caio Vaz Rimoli (1) , Frank Schnorrer (4) , Francois Robin (7) , Andrew Wilde (3) , Loic Legoff (1) , Jean-Denis Pedelacq (8) , Stephanie Cabantous (9) , Sergio Rincon (5) , Cristel Chandre (10) , Sophie Brasselet (1) , Manos Mavrakis (1)

1 MOSAIC - MOSAIC
2 IBPS - Institut de Biologie Paris Seine
3 University of Toronto
4 IBDM - Institut de Biologie du Développement de Marseille
5 IBFG - Instituto de Biología Funcional y Genómica [Salamanca, Spain]
6 Biologie Cellulaire et Cancer
7 LBD-E17 - Dynamique de l'actomyosine corticale lors du développement et de la morphogenèse = Cortical actomyosin dynamics in development and morphogenesis
8 IPBS - Institut de pharmacologie et de biologie structurale
9 CRCT - Centre de Recherches en Cancérologie de Toulouse
10 I2M - Institut de Mathématiques de Marseille

The cytoskeletal protein actin is crucial for cell shape and integrity throughout eukaryotes. Actin filaments perform essential biological functions, including muscle contraction, cell division and tissue morphogenesis. These diverse activities are achieved through the ability of actin filaments to be arranged into diverse architectures, but a detailed appreciation of the dynamic organizational state of the actin filaments has been hindered by available tools. Fluorescence polarization microscopy is uniquely placed for measuring actin organization by exploiting the sensitivity of polarized light excitation to the orientation of fluorophores attached to actin filaments. By engineering constrained fluorescent protein fusions to widely used actin localization reporters, we have succeeded in developing novel genetically-encoded reporters for non-invasive, quantitative measurements of actin filament organization in living cells by fluorescence polarization microscopy. We show examples of actin organization measurements in living mammalian cells in culture, as well as in vivo in fission yeast, C. elegans and Drosophila.

Mots clés: actin filaments; molecular organization; molecular order; molecular orientation; filament alignment; fluorescence polarization microscopy; morphogenesis; GFP mobility

Domaines: Biologie structurale [q-bio.BM]; Organisation et fonctions cellulaires [q-bio.SC]; Biotechnologies

Carla Silva Martins, Francois Iv, Shashi Kumar Suman, Thomas Panagiotou, Clara Sidor, et al.. Genetically encoded reporters of actin filament organization in living cells and tissues. 2024. ⟨hal-04563732⟩