Androgen receptor (AR) exerts its diverse biological functions primarily through its ability to regulate gene expression. As a member of nuclear receptor superfamily, AR recruits various coactivators to facilitate its transcriptional activity. The ligand-binding domains (LBD) of AR is believed to play a role in coactivator recruitment through a direct interaction between a hydrophobic coactivator binding groove in the LBD and a FXXLF or LXXLL motif within coactivators. In this study, we provide multiple lines of evidence showing that the FXXLF motif-containing ARA70 and ARA54 exhibit strong hormone-dependent interaction with the AR LBD but poorly with full-length AR. This drastic difference in interaction with ARA70 and ARA54 between the AR LBD and full-length AR is due to the hormone-dependent N-C interaction of AR. Like the AR LBD, full-length AR mutants defective in the N-C interaction exhibit strong hormone-dependent interaction with ARA70 and ARA54. Thus, our results suggest that in the full-length context the hydrophobic coactivator binding groove in the LBD is normally engaged in the liganded induced AR N-C interaction and thus restricted from interaction with other proteins. This finding raises fundamental question as to how AR recruits coactivators to regulate gene transcription.