β1-adrenergic receptor (β1AR) is a potent regulator of cardiac function. Previously, we showed that recycling endosome (RE) is key organelle of the intracellular trafficking of β1AR induced by ligand stimulus. Here, we found that HACE1, an E3 ligase, is localized in RE and ubiquitinates β1AR internalized by the ligand isoproterenol (Iso). Silencing of HACE1 by siRNA resulted in a decline of β1AR on the cell surface at steady state. Given that the ubiquitinated Rab11 functions as a recycling factor, ubiquitinated Rab11 mediated by HACE1 might recycle β1AR to the cell surface. Ubiquitination of β1AR was increased by Iso treatment, which was further enhanced with either bafilomycin A1(BafA1), a lysosome inhibitor or MG132, a proteasome inhibitor. β1AR was co-localized with Rab12 at RE and lysosome and interacted with each other. The co-localization at lysosome was strengthened in the presence of BafA1, and the interaction was enhanced by Iso stimulus. Silencing of Rab12 led to stabilization of β1AR with or without Iso. These data suggest that Rab11 and Rab12 were involved in the recycling and degradation of β1AR through ubiquitination of Rab11and β1AR mediated by HACE1 in RE, respectively.