A proteomic approach to identify endosomal cargoes controlling cancer invasiveness

Diaz-Vera, J., Palmer, S., Hernandez-Fernaud, J. R., Dornier, E., Mitchell, L. E., MacPherson, I. , Edwards, J. , Zanivan, S. and Norman, J. C. (2017) A proteomic approach to identify endosomal cargoes controlling cancer invasiveness. Journal of Cell Science, 130(4), pp. 697-711. (doi: 10.1242/jcs.190835) (PMID:28062852)

Abstract

We have previously shown that Rab17 - a small GTPase associated with epithelial polarity - is specifically suppressed by ERK2 signalling to promote an invasive phenotype. However, the mechanisms through which Rab17 loss permits invasiveness, and the endosomal cargoes that are responsible for mediating this are not known. Using quantitative mass spectrometry-based proteomics, we have found that knockdown of Rab17 leads to highly selective reduction in the cellular levels of a v-SNARE (Vamp8). Moreover, proteomics and immunofluorescence indicate that Vamp-8 is associated with Rab17 at late endosomes. Reduced levels of Vamp8 promote transition between ductal carcinoma in situ (DCIS) and a more invasive phenotype. We developed an unbiased proteomic approach to elucidate the complement of receptors that redistributes between endosomes and the plasma membrane, and have pin-pointed neuropilin-2 (NRP2) as a key pro-invasive cargo of Rab17/Vamp8-regulated trafficking. Indeed, reduced Rab17 or Vamp8 levels lead to increased mobilisation of NRP2-containing late endosomes and upregulated cell surface expression of NRP2. Finally, we show that NRP2 is required for the basement membrane disruption which accompanies transition between DCIS and a more invasive phenotype.

• Item Type: Articles
• Additional Information: Supported by Cancer Research UK (grant numbers; JCN, C596/A18277 and C596/A17196; SZ, C596/A12935), and Breast Cancer Now (grant to JCN, IM and ED).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Zanivan, Professor Sara and Dornier, Mr Emmanuel Greg and Edwards, Professor Joanne and MacPherson, Professor Iain and Norman, Professor James and Mitchell, Mrs Louise
• Authors: Diaz-Vera, J., Palmer, S., Hernandez-Fernaud, J. R., Dornier, E., Mitchell, L. E., MacPherson, I., Edwards, J., Zanivan, S., and Norman, J. C.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Journal of Cell Science
• Publisher: Company of Biologists
• ISSN: 0021-9533
• ISSN (Online): 1477-9137
• Published Online: 05 January 2017
• Copyright Holders: Copyright © 2017 The Authors
• First Published: First published in Journal of Cell Science 130(4):697-711
• Publisher Policy: Reproduced under a Creative Commons License