Info

Proteins N expressed:

Proteins N expressed:

ß-Galactosidase

Glucocorticoid receptor

GR ligand-binding domain

ß-Galactosidase

Glucocorticoid receptor

GR ligand-binding domain

▲ EXPERIMENTAL FIGURE 11-43 Fusion proteins from expression vectors demonstrate that the hormone-binding domain of the glucocorticoid receptor (GR) mediates translocation to the nucleus in the presence of hormone. Cultured animal cells were transfected with expression vectors encoding the proteins diagrammed at the bottom. Immunofluorescence with a labeled antibody specific for p-galactosidase was used to detect the expressed proteins in transfected cells. (a) In cells that expressed p-galactosidase alone, the enzyme was localized to the cytoplasm In the presence and absence of the glucocorticoid hormone dexamethasone (Dex). (b) In cells that expressed a fusion protein consisting of ß-galactosidase and the entire glucocorticoid receptor (GR), the fusion protein was present in the cytoplasm in the absence of hormone but was transported to the nucleus in the presence of hormone. (c) Cells that expressed a fusion protein composed of ß-galactosidase and just the GR ligand-binding domain (light purple) also exhibited hormone-dependent transport of the fusion protein to the nucleus. [From D. Picard and K. R. Yamamoto, 1987, EMBO J. 6:3333; courtesy of the authors.]

Hormone

Hormone

▲ FIGURE 11-44 Model of hormone-dependent gene activation by a homodimeric nuclear receptor. In the absence of hormone, the receptor is kept in the cytoplasm by interaction between its ligand-binding domain (LBD) and inhibitor proteins. When hormone is present, it diffuses through the plasma membrane and binds to the ligand-binding domain, causing a conformational change that releases the receptor from the inhibitor proteins. The receptor with bound ligand is then translocated into the nucleus, where its DNA-binding domain (DBD) binds to response elements, allowing the ligand-binding domain and an additional activation domain (AD) at the N-terminus to stimulate transcription of target genes.

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