Prenylation-dependent association of protein-tyrosine phosphatases PRL-1, -2, and -3 with the plasma membrane and the early endosome

J Biol Chem. 2000 Jul 14;275(28):21444-52. doi: 10.1074/jbc.M000453200.

Abstract

PRL-1, -2, and -3 represent a novel class of protein-tyrosine phosphatase with a C-terminal prenylation motif. Although PRL-1 has been suggested to be associated with the nucleus, the presence of three highly homologous members and the existence of a prenylation motif call for a more detailed examination of their subcellular localization. In the present study, we first demonstrate that mouse PRL-1, -2, and -3 are indeed prenylated. Examination of N-terminal epitope-tagged PRL-1, -2, and -3 expressed in transiently transfected cells suggests that PRL-1, -2, and -3 are present on the plasma membrane and intracellular punctate structures. Stable Chinese hamster ovary cells expressing PRL-1 and -3 in an inducible manner were established. When cells were treated with brefeldin A, PRL-1 and -3 accumulated in a collapsed compact structure around the microtubule-organizing center. Furthermore, PRL-1 and -3 redistributed into swollen vacuole-like structures when cells were treated with wortmannin. These characteristics of PRL-1 and -3 are typical for endosomal proteins. Electron microscope immunogold labeling reveals that PRL-1 and -3 are indeed associated with the plasma membrane and the early endosomal compartment. Expression of PRL-3 is detected in the epithelial cells of the small intestine, where PRL-3 is present in punctate structures in the cytoplasm. When cells are treated with FTI-277, a selective farnesyltransferase inhibitor, PRL-1, -2, and -3 shifted into the nucleus. Furthermore, a mutant form of PRL-2 lacking the C-terminal prenylation signal is associated with the nucleus. These results establish that the primary association of PRL-1, -2, and -3 with the membrane of the cell surface and the early endosome is dependent on their prenylation and that nuclear localization of these proteins may be triggered by a regulatory event that inhibits their prenylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkyl and Aryl Transferases / antagonists & inhibitors
  • Animals
  • Brefeldin A / pharmacology
  • CHO Cells
  • Cell Membrane / enzymology*
  • Cell Membrane / ultrastructure
  • Cricetinae
  • Endosomes / drug effects
  • Endosomes / enzymology*
  • Endosomes / ultrastructure
  • Enzyme Inhibitors / pharmacology
  • Farnesyltranstransferase
  • Immediate-Early Proteins / metabolism*
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / enzymology
  • Methionine / analogs & derivatives
  • Methionine / pharmacology
  • Mice
  • Microvilli / enzymology
  • Microvilli / ultrastructure
  • Protein Prenylation
  • Protein Tyrosine Phosphatases / metabolism*
  • Recombinant Proteins / metabolism
  • Transfection

Substances

  • Enzyme Inhibitors
  • FTI 277
  • Immediate-Early Proteins
  • Ptp4a3 protein, mouse
  • Recombinant Proteins
  • Brefeldin A
  • Methionine
  • Alkyl and Aryl Transferases
  • Farnesyltranstransferase
  • Protein Tyrosine Phosphatases
  • Ptp4a1 protein, mouse
  • Ptp4a2 protein, mouse