c-Jun potentiates the functional interaction between the amino and carboxyl termini of the androgen receptor.
The transactivation functions of the human androgen receptor (hAR) are regulated by several accessory factors that can be either positive or negative. One factor that has been previously shown to mediate hAR transactivation is the proto-oncoprotein c-Jun. The positive effect is a primary one, can be exerted by both endogenous and exogenous c-Jun, and requires multiple regions of c-Jun. However, the exact mechanism by which c-Jun exerts its enhancing function is unknown. In this study, we have used a mammalian two-hybrid system to ask if c-Jun influences the ligand-dependent amino- to carboxyl-terminal (N-to-C) interaction of hAR, which is thought to be responsible for the homodimerization of this receptor. Our results show that c-Jun enhances both hAR N-to-C terminal interaction and DNA binding in vitro. We have also tested a panel of c-Jun and c-Fos mutants for their activities on the N-to-C interaction, and the data demonstrate that the activities of these mutants parallel their activities on hAR transactivation. A mutation in the hAR activation function-2 (AF-2) abrogates N-to-C interaction, DNA binding, and transactivation, and these activities are not rescued by exogenous c-Jun. Interestingly, the p160 coactivator TIF2 can stimulate hAR N-to-C interaction, a finding consistent with the effect on hAR transactivation. These data strongly suggest that the hAR N-to-C interaction is the target of c-Jun action, and this activity requires a functional receptor AF-2.
Bubulya
A
A
Department of Biological Sciences, University of Toledo, Toledo, Ohio 43606, USA.
Chen
S Y
SY
Fisher
C J
CJ
Zheng
Z
Z
Shen
X Q
XQ
Shemshedini
L
L
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D017930:Genes, Reporter; D016758:Genes, jun / Q000235:genetics; D006801:Humans; D008024:Ligands; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism* / Q000502:physiology*; D015533:Transcriptional Activation; D014162:Transfection; D020798:Two-Hybrid System Techniques
D000596:Amino Acids; D008024:Ligands; D016755:Proto-Oncogene Proteins c-jun; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase