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DMSO Enhances TGF‐β Activity by Recruiting the Type II TGF‐β Receptor From Intracellular Vesicles to the Plasma Membrane
Author(s) -
Huang Shuan Shian,
Chen ChunLin,
Huang Franklin W.,
Hou WeiHsien,
Huang Jung San
Publication year - 2016
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25448
Subject(s) - vesicle , intracellular , western blot , microbiology and biotechnology , cytoplasm , chemistry , transfection , lipid raft , exosome , receptor , biology , biochemistry , microvesicles , membrane , gene , microrna
Dimethyl sulfoxide (DMSO) is used to treat many diseases/symptoms. The molecular basis of the pharmacological actions of DMSO has been unclear. We hypothesized that DMSO exerts some of these actions by enhancing TGF‐β activity. Here we show that DMSO enhances TGF‐β activity by ∼3–4‐fold in Mv1Lu and NMuMG cells expressing Smad‐dependent luciferase reporters. In Mv1Lu cells, DMSO enhances TGF‐β‐stimulated expression of P‐Smad2 and PAI‐1. It increases cell‐surface expression of TGF‐β receptors (TβR‐I and/or TβR‐II) by ∼3–4‐fold without altering their cellular levels as determined by 125 I‐labeled TGF‐β‐cross‐linking/Western blot analysis, suggesting the presence of large intracellular pools in these cells. Sucrose density gradient ultracentrifugation/Western blot analysis reveals that DMSO induces recruitment of TβR‐II (but not TβR‐I) from its intracellular pool to plasma‐membrane microdomains. It induces more recruitment of TβR‐II to non‐lipid raft microdomains than to lipid rafts/caveolae. Mv1Lu cells transiently transfected with TβR‐II‐HA plasmid were treated with DMSO and analyzed by indirect immunofluoresence staining using anti‐HA antibody. In these cells, TβR‐II‐HA is present as a vesicle‐like network in the cytoplasm as well as in the plasma membrane. DMSO causes depletion of TβR‐II‐HA‐containing vesicles from the cytoplasm and co‐localization of TβR‐II‐HA and cveolin‐1 at the plasma membrane. These results suggest that DMSO, a fusogenic substance, enhances TGF‐β activity presumably by inducing fusion of cytoplasmic vesicles (containing TβR‐II) and the plasma membrane, resulting in increased localization of TβR‐II to non‐lipid raft microdomains where canonical signaling occurs. Fusogenic activity of DMSO may play a pivotal role in its pharmacological actions involving membrane proteins with large cytoplasmic pools. J. Cell. Biochem. 117: 1568–1579, 2016. © 2015 Wiley Periodicals, Inc.