Predominant Intracellular Localization of the Type I Transforming Growth Factor-β Receptor and Increased Nuclear Accumulation after Growth Arrest

• Published in: Experimental cell research 2000-07, Vol.258 (1), p.121-134
• Main Authors: Zwaagstra, John C., Guimond, Alain, O'Connor-McCourt, Maureen D.
• Format: Article
• Language: English
• Subjects: Activin Receptors, Type I; Animals; Cell Cycle - drug effects; Cell Cycle - physiology; Cell Line; Cell Membrane - physiology; Cell Nucleus - physiology; Cyclin-Dependent Kinases - antagonists & inhibitors; Endoplasmic Reticulum - physiology; Enzyme Inhibitors - pharmacology; Gene Expression Regulation - drug effects; Golgi Apparatus - physiology; Green Fluorescent Proteins; growth arrest; Humans; intracellular; Kinetin; Luminescent Proteins - analysis; Luminescent Proteins - genetics; Lung Neoplasms; Mink; nucleus; Protein-Serine-Threonine Kinases - analysis; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - physiology; Purines - pharmacology; Receptors, Transforming Growth Factor beta - analysis; Receptors, Transforming Growth Factor beta - genetics; Receptors, Transforming Growth Factor beta - physiology; Recombinant Fusion Proteins - biosynthesis; Respiratory Mucosa; Signal Transduction - drug effects; Signal Transduction - physiology; TGF-β receptor; Transfection; Transforming Growth Factor beta - pharmacology; Tumor Cells, Cultured
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1006/excr.2000.4905

Transforming growth factor-β (TGF-β) signaling requires the functional interaction of two distinct receptors, type I (RI) and type II (RII), at the cell surface. Exposure of cells to TGF-β results in receptor internalization and down-regulation (Zwaagstra et al., 1999, Exp. Cell Res. 252, 352–362); however, little is known about the subsequent fate of RI or RII. In this study the cellular distribution of RI was examined in cells before and after treatment with ligand. RI was localized by immunocytochemistry and confocal microscopy using two polyclonal antisera directed against two different epitopes, one in the C-terminal region and one in the N-terminal region of the cytoplasmic domain. The majority of RI molecules in untreated Mv1Lu and A549 cells were found to be intracellular. Treatment of Mv1Lu and A549 cells with 100 pM TGF-β1 for 24 h at 37°C caused a redistribution of surface RI on Mv1Lu cells, as evidenced by surface RI aggregation. Unexpectedly, this TGF-β1 treatment also caused redistribution and accumulation of intracellular RI in and around the nucleus for both Mv1Lu and A549 cells. Nuclear accumulation of RI was also promoted independently of ligand receptor activation by treatment of Mv1Lu cells with olomoucine, an agent that results in growth arrest. The capacity of RI to localize in the nucleus was confirmed by microscopic examination of 293 cells transiently expressing RI fused to green fluorescent protein (RI-GFP). Olomoucine treatment of these cells resulted in the movement of RI-GFP into the nucleus. Our results indicate that growth arrest alters intracellular transport/routing of RI and may indicate that RI functions not only at the cell surface but inside the cell as well.