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Mediator subunit MED1 modulates intranuclear dynamics of the thyroid hormone receptor
Author(s) -
Femia Matthew R.,
Evans Rochelle M.,
Zhang Jibo,
Sun Xiaopeng,
Lebegue Caroline J.,
Roggero Vincent R.,
Allison Lizabeth A.
Publication year - 2020
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.29532
Subject(s) - transactivation , mediator , fluorescence recovery after photobleaching , nuclear export signal , thyroid hormone receptor , nuclear receptor , biology , population , cell nucleus , cytosol , microbiology and biotechnology , nuclear localization sequence , nuclear transport , protein subunit , medicine , endocrinology , transcription factor , hormone , cytoplasm , gene , biochemistry , environmental health , membrane , enzyme
The thyroid hormone receptors (TRs) mediate thyroid hormone (T 3 )‐dependent gene expression. The nuclear import and export signals that direct TR shuttling are well characterized, but little is known about factors modulating nuclear retention. We used fluorescence‐based nucleocytoplasmic scoring and fluorescence recovery after photobleaching in transfected cells to investigate whether Mediator subunits MED1 and MED13 play a role in nuclear retention of TR. When MED1 was overexpressed, there was a striking shift towards a greater nuclear localization of TRβ1 and the oncoprotein v‐ErbA, subtypes with cytosolic populations at steady‐state, and TRβ1 intranuclear mobility was reduced. For TRα1, there was no observable change in its predominantly nuclear distribution pattern or mobility. Consistent with a role for MED1 in nuclear retention, the cytosolic TRα1 and TRβ1 population were significantly greater in MED1 −/− cells, compared with MED1 +/+ cells. Exposure to T 3 and epidermal growth factor, which induces MED1 phosphorylation, also altered TR intranuclear dynamics. Overexpression of miR‐208a, which downregulates MED13, led to a more cytosolic distribution of nuclear‐localized TRα1; however, overexpression of MED13 had no effect on TRβ1 localization. The known binding site of MED1 overlaps with a transactivation domain and nuclear export signal in helix 12 of TR's ligand‐binding domain (LBD). Coimmunoprecipitation assays demonstrated that TR's LBD interacts directly with exportins 5 and 7, suggesting that binding of exportins and MED1 to TR may be mutually exclusive. Collectively, our data provide evidence that MED1 promotes nuclear retention of TR, and highlight the dual functionality of helix 12 in TR transactivation and nuclear export.