Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells

Motoaki Seki; Akira Watanabe; Satoshi Enomoto; Takeshi Kawamura; Hirotaka Ito; Tatsuhiko Kodama; Takao Hamakubo; Hiroyuki Aburatani

doi:10.1016/j.febslet.2010.05.009

Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells

FEBS Lett. 2010 Jul 2;584(13):2909-15. doi: 10.1016/j.febslet.2010.05.009. Epub 2010 May 13.

Authors

Motoaki Seki¹, Akira Watanabe, Satoshi Enomoto, Takeshi Kawamura, Hirotaka Ito, Tatsuhiko Kodama, Takao Hamakubo, Hiroyuki Aburatani

Affiliation

¹ Genome Science Division, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.

PMID: 20471383
DOI: 10.1016/j.febslet.2010.05.009

Abstract

ROBO1 is a receptor mediating Slit-induced repulsive action on axon guidance and differentially expressed in human cancers. Although ROBO1 ectodomain has been detected, the cleavage site had not been determined. In this study we identified the precise cleavage site of ROBO1. We also report multi-step proteolysis of ROBO1 by metalloproteinases and gamma-secretase, producing two carboxy-terminal fragments, ROBO1-CTF1 at 129-kDa and ROBO1-CTF2 at 118-kDa. We have further demonstrated nuclear accumulation of ROBO1, which is abolished by either a metalloproteinase inhibitor TAPI-1 or a gamma-secretase inhibitor L-685,458. ROBO1 may function beyond the receptor through stepwise cleavages and translocation to the nucleus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Amyloid Precursor Protein Secretases / antagonists & inhibitors
Amyloid Precursor Protein Secretases / metabolism*
Carbamates / pharmacology
Cell Nucleus / metabolism*
Dipeptides / pharmacology
Hep G2 Cells
Humans
Hydroxamic Acids / pharmacology
Immunoblotting
Immunohistochemistry
Leupeptins / pharmacology
Metalloproteases / antagonists & inhibitors
Metalloproteases / metabolism*
Molecular Sequence Data
Nerve Tissue Proteins / chemistry
Nerve Tissue Proteins / metabolism*
Receptors, Immunologic / chemistry
Receptors, Immunologic / metabolism*
Roundabout Proteins
Sequence Homology, Amino Acid
Tandem Mass Spectrometry

Substances

Carbamates
Dipeptides
Hydroxamic Acids
L 685458
Leupeptins
N-((2-(hydroxyaminocarbonyl)methyl)-4-methylpentanoyl)-3-(2'-naphthyl)alanylalanine, 2-aminoethylamide
N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide
Nerve Tissue Proteins
Receptors, Immunologic
Amyloid Precursor Protein Secretases
Metalloproteases
benzyloxycarbonylleucyl-leucyl-leucine aldehyde