Interaction of the Ubc9 human homologue with c-Jun and with the glucocorticoid receptor.
Glucocorticoid hormones convert the glucocorticoid receptor (GR) from an inactive cytosolic complex to a nuclear form that regulates transcription. Binding of GR to palindromic DNA-recognition sites (hormone response elements) leads to activated target gene transcription. GR also exerts negative actions on transcription, e.g., by interfering with the function of several other transcription factors such as AP-1, NK-kappa B, CREB, and Oct-1. Physical interactions of GR with AP-1 subunits are readily detectable but do not seem sufficient since nonrepressing GR mutants still interact in vitro, so that specific conformational changes and/or interactions with additional partner proteins may be required for negative action. In an attempt to find such partner proteins, we defined regions of c-Jun and GR essential for mutual interference and used in those a yeast two-hybrid screen for interacting proteins. Repeatedly we isolated overlapping cDNA sequences of one protein interaction with both c-Jun and GR. This protein does not interact with c-Fos or a non-repressing GR mutant and expressed in mammalian cells does not substantially affect AP-1 or GR activity. Interestingly, however, the protein rescues yeast cells from the toxic effects of the GR fragment used for screening. The protein represents the human homologue of the yeast E2 ubiquitin-conjugating enzyme, Ubc9; its specific interactions with both GR and c-Jun, but not mutant GR, suggest that it may exert physiologic regulatory functions.
Göttlicher
M
M
Forschungzentrum Karlsruhe, Institute of Genetics, Germany.
Heck
S
S
Doucas
V
V
Wade
E
E
Kullmann
M
M
Cato
A C
AC
Evans
R M
RM
Herrlich
P
P
D000818:Animals; D001665:Binding Sites; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D005804:Genes, Lethal; D006801:Humans; D006822:Hybrid Cells; D008025:Ligases / Q000737:chemistry / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D005656:Fungal Proteins; D016755:Proto-Oncogene Proteins c-jun; D011965:Receptors, Glucocorticoid; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D044763:Ubiquitin-Conjugating Enzymes; D008025:Ligases; D000098843:Ubiquitin-Conjugating Enzyme UBC9