Format

Send to

Choose Destination
J Cell Sci. 2017 Feb 15;130(4):697-711. doi: 10.1242/jcs.190835. Epub 2017 Jan 6.

A proteomic approach to identify endosomal cargoes controlling cancer invasiveness.

Author information

1
Beatson Institute for Cancer Research, Garscube Estate, Glasgow G61 1BD, UK.
2
Wolfson Wohl Cancer Research Centre, Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK.
3
Beatson Institute for Cancer Research, Garscube Estate, Glasgow G61 1BD, UK s.zanivan@beatson.gla.ac.uk j.norman@beatson.gla.ac.uk.
4
Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK.

Abstract

We have previously shown that Rab17, a small GTPase associated with epithelial polarity, is specifically suppressed by ERK2 (also known as MAPK1) 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 unknown. Using quantitative mass spectrometry-based proteomics, we have found that knockdown of Rab17 leads to a highly selective reduction in the cellular levels of a v-SNARE (Vamp8). Moreover, proteomics and immunofluorescence indicate that Vamp8 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- and 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 that accompanies the transition between DCIS and a more invasive phenotype.

KEYWORDS:

Breast cancer; Cell migration; Ductal carcinoma in situ; Invasion; Invasive ductal carcinoma; Late endosome; Neuropilin-2; Rab17; SILAC; Vamp8

PMID:
28062852
PMCID:
PMC5339883
DOI:
10.1242/jcs.190835
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center