Functional and Genetic Analysis of VAB-10 Spectraplakin in Caenorhabditis elegans.
Fiche publication
Date publication
janvier 2016
Journal
Methods in enzymology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr GALLY Christelle
Tous les auteurs :
Gally C, Zhang H, Labouesse M
Lien Pubmed
Résumé
Intermediate filaments (IFs) are involved in multiple cellular processes that are essential for the maintenance of cell and tissue integrity. To achieve this crucial function, IFs have to be organized as long and resistant filaments across the cells and to be tightly anchored at the cell periphery. This anchoring takes place at the level desmosomes and hemidesmosomes through proteins belonging to the spectraplakin family. Here, we focus on the sole nematode Caenorhabditis elegans spectraplakin locus vab-10 that is essential to connect the epidermis to the cuticle apically and to the muscles basally. After briefly reviewing the structure of the gene, we first present the genetic tools available to study this gene as well as the reagents to examine the distribution of its translation products. We discuss the functional assays that enable examining their function. Finally, we detail a genetic method to identify spectraplakin functional partners through RNAi screens, and a biochemical method to examine the phosphorylation status of IFs.
Mots clés
Animals, Base Sequence, Caenorhabditis elegans, cytology, Caenorhabditis elegans Proteins, chemistry, Exons, Mutation, Protein Structure, Tertiary
Référence
Meth. Enzymol.. 2016 ;569:407-30