1. Tks5 SH3 domains exhibit differential effects on invadopodia development.
- Author
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Daly C, Logan B, Breeyear J, Whitaker K, Ahmed M, and Seals DF
- Subjects
- Adaptor Proteins, Vesicular Transport chemistry, Carcinoma metabolism, Carcinoma pathology, Cell Line, Tumor, Cell Movement genetics, Cortactin genetics, Fibroblasts metabolism, Fibroblasts pathology, Gelatin genetics, Gelatin metabolism, Gene Expression Regulation, Neoplastic genetics, Humans, Male, Mutation genetics, Phosphorylation, Podosomes genetics, Prostatic Neoplasms metabolism, Prostatic Neoplasms pathology, Protein Interaction Domains and Motifs genetics, src Homology Domains genetics, Adaptor Proteins, Vesicular Transport genetics, Carcinoma genetics, Prostatic Neoplasms genetics, src-Family Kinases genetics
- Abstract
The Src substrate Tks5 helps scaffold matrix-remodeling invadopodia in invasive cancer cells. Focus was directed here on how the five SH3 domains of Tks5 impact that activity. Mutations designed to inhibit protein-protein interactions were created in the individual SH3 domains of Tks5, and the constructs were introduced into the LNCaP prostate carcinoma cell line, a model system with intrinsically low Tks5 expression and which our lab had previously showed the dependence of Src-dependent Tks5 phosphorylation on invadopodia development. In LNCaP cells, acute increases in wild-type Tks5 led to increased gelatin matrix degradation. A similar result was observed when Tks5 was mutated in its 4th or 5th SH3 domains. This was in contrast to the 1st, 2nd, and 3rd SH3 domain mutations of Tks5 where each had a remarkable accentuating effect on gelatin degradation. Conversely, in the invadopodia-competent Src-3T3 model system, mutations in any one of the first three SH3 domains had a dominant negative effect that largely eliminated the presence of invadopodia, inhibited gelatin degradation activity, and redistributed both Src, cortactin, and Tks5 to what are likely endosomal compartments. A hypothesis involving Tks5 conformational states and the regulation of endosomal trafficking is presented as an explanation for these seemingly disparate results., Competing Interests: The authors have declared that no competing interests exist. more...
- Published
- 2020
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