Combining laser capture microdissection and proteomics reveals an active translation machinery controlling invadosome formation

Zakaria Ezzoukhry, Elodie Henriet, Fabrice P. Cordelières, Jean-William Dupuy, Marlène Maître, Nathan Gay, Sylvaine Di-Tommaso, Luc Mercier, Jacky G. Goetz, Marion Peter, Frédéric Bard, Violaine Moreau, Anne-Aurélie Raymond and Frédéric Saltel ()
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Zakaria Ezzoukhry: BaRITOn Bordeaux Research in Translational Oncology
Elodie Henriet: BaRITOn Bordeaux Research in Translational Oncology
Fabrice P. Cordelières: Université de Bordeaux
Jean-William Dupuy: Université de Bordeaux
Marlène Maître: INSERM
Nathan Gay: BaRITOn Bordeaux Research in Translational Oncology
Sylvaine Di-Tommaso: BaRITOn Bordeaux Research in Translational Oncology
Luc Mercier: 1 Place de L’Hôpital
Jacky G. Goetz: 1 Place de L’Hôpital
Marion Peter: CNRS, Univ. Montpellier
Frédéric Bard: Institute of Molecular and Cell Biology
Violaine Moreau: BaRITOn Bordeaux Research in Translational Oncology
Anne-Aurélie Raymond: BaRITOn Bordeaux Research in Translational Oncology
Frédéric Saltel: BaRITOn Bordeaux Research in Translational Oncology

Nature Communications, 2018, vol. 9, issue 1, 1-11

Abstract: Abstract Invadosomes are F-actin-based structures involved in extracellular matrix degradation, cell invasion, and metastasis formation. Analyzing their proteome is crucial to decipher their molecular composition, to understand their mechanisms, and to find specific elements to target them. However, the specific analysis of invadosomes is challenging, because it is difficult to maintain their integrity during isolation. In addition, classical purification methods often suffer from contaminations, which may impair data validation. To ensure the specific identification of invadosome components, we here develop a method that combines laser microdissection and mass spectrometry, enabling the analysis of subcellular structures in their native state based on low amounts of input material. Using this combinatorial method, we show that invadosomes contain specific components of the translational machinery, in addition to known marker proteins. Moreover, functional validation reveals that protein translation activity is an inherent property of invadosomes, which is required to maintain invadosome structure and activity.

Date: 2018
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DOI: 10.1038/s41467-018-04461-9

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