ARF6-JIP3/4 regulate endosomal tubules for MT1-MMP exocytosis in cancer invasion

J Cell Biol. 2015 Oct 26;211(2):339-58. doi: 10.1083/jcb.201506002.

Abstract

Invasion of cancer cells into collagen-rich extracellular matrix requires membrane-tethered membrane type 1-matrix metalloproteinase (MT1-MMP) as the key protease for collagen breakdown. Understanding how MT1-MMP is delivered to the surface of tumor cells is essential for cancer cell biology. In this study, we identify ARF6 together with c-Jun NH2-terminal kinase-interacting protein 3 and 4 (JIP3 and JIP4) effectors as critical regulators of this process. Silencing ARF6 or JIP3/JIP4 in breast tumor cells results in MT1-MMP endosome mispositioning and reduces MT1-MMP exocytosis and tumor cell invasion. JIPs are recruited by Wiskott-Aldrich syndrome protein and scar homologue (WASH) on MT1-MMP endosomes on which they recruit dynein-dynactin and kinesin-1. The interaction of plasma membrane ARF6 with endosomal JIPs coordinates dynactin-dynein and kinesin-1 activity in a tug-of-war mechanism, leading to MT1-MMP endosome tubulation and exocytosis. In addition, we find that ARF6, MT1-MMP, and kinesin-1 are up-regulated in high-grade triple-negative breast cancers. These data identify a critical ARF6-JIP-MT1-MMP-dynein-dynactin-kinesin-1 axis promoting an invasive phenotype of breast cancer cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-Ribosylation Factors / genetics
  • ADP-Ribosylation Factors / metabolism*
  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Collagen / metabolism
  • Endosomes / metabolism
  • Exocytosis / physiology
  • Female
  • HEK293 Cells
  • Humans
  • Kinesin / metabolism
  • Matrix Metalloproteinase 14 / genetics
  • Matrix Metalloproteinase 14 / metabolism*
  • Microfilament Proteins / metabolism
  • Neoplasm Invasiveness
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Protein Transport
  • RNA Interference
  • RNA, Small Interfering
  • Spheroids, Cellular
  • Triple Negative Breast Neoplasms / pathology*
  • Tumor Cells, Cultured

Substances

  • Adaptor Proteins, Signal Transducing
  • KIF3A protein, human
  • KIF5B protein, human
  • MAPK8IP3 protein, human
  • Microfilament Proteins
  • Nerve Tissue Proteins
  • RNA, Small Interfering
  • SPAG9 protein, human
  • WASH protein, human
  • Collagen
  • MMP14 protein, human
  • Matrix Metalloproteinase 14
  • Kinesin
  • ADP-Ribosylation Factors
  • ADP-ribosylation factor 6