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Olivier Destaing — Moving from genetic to optogenetics : a new way to understand the complex dynamics of invadosomes
Speaker :
Olivier Destaing, Institut Albert Bonniot INSERM U823, Equipe 1 DySAD ERL CNRS 3148, Grenoble
When :
Wednesday 23 May at 11h
Where :
Salle 116
Title :
Moving from genetic to optogenetics : a new way to understand the complex dynamics of invadosomes
Abstract :
Invadosomes, comprising invadopodia and podosomes, are very dynamics adhesion structures involved in tissue invasion, characterized by an intense actin polymerization/depolymerization associated with β1 and β3 integrins and coupled to extracellular matrix degradation (ECM) activity. In order to explore the molecular mechanisms controlling this machinery, we induced the formation of invadosomes by expressing the constitutive active form of Src, SrcYF, in different MEF cells knock-out for some specific targets. Using β1-/- or β3-/- cells, it appeared that β1A but not β3 integrins are surprisingly essential for initiation of invadosome formation. In parallel, we also determined that the large GTPase dynamin (playing a key role in endocytosis) is also essential for actin assembly of invadosomes. The only limitation of genetic approach is its poor temporal resolution. There is indeed a large time difference between the depletion of a specific protein (days) and the characteristic time of the observed structures (min). In order to determine direct effect of integrin and dynamin depletion on invadosome, an optogenetic approach based on the photosensitizer KillerRed was developed to obtain loss of function at the minute scale. By affecting these molecules, we are even able to disorganize invadosomes without affecting focal adhesions.
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