Many important physiological roles of the urocortin (UCN)-family of peptides as well as corticotropin-releasing hormone (CRH), involve the type 2 CRH receptor (CRH-R2) and downstream activation of multiple pathways. To characterize molecular determinants of CRH-R2 functional activity, we used HEK293 cells overexpressing recombinant CRH-R2 and investigated mechanisms involved in attenuation of CRH-R2 signaling activity and uncoupling from intracellular effectors. CRH-R2-mediated adenylyl cyclase activation was sensitive to homologous desensitization induced by pretreatment with either UCN-II or the weaker agonist CRH. CRH-R2 activation induced transient -arrestin1 and -arrestin2, as well as clathrin, recruitment to the plasma membrane. -Arrestin2 appeared to be the main -arrestin subtype associated with the receptor. This was followed by CRH-R2 endocytosis in a mechanism that exhibited distinct agonist-dependent temporal characteristics. CRH-R2 also induced transient activation of the ERK1/2 and p38MAPK signaling cascades that peaked at 5min and returned to basal within 20–30min. Unlike p38MAPK, activated ERK1/2 was localized both in the cytoplasm and nucleus. Experiments employing inhibitors of receptor endocytosis showed that CRH-R2-MAPK interaction does not require -arrestin, clathrin or receptor endocytosis. Site-directed mutagenesis studies on CRH-R2 C-terminus showed that the aminoacid cassette TAAV at the end of the C-terminus is important for CRH-R2 signaling since loss of a potential phospho-acceptor site in mutant receptors containing deletion or Ala substitution of the cassette TAAV resulted in reduced ERK1/2 activation and accelerated receptor internalization. These findings provide new insights about the signaling mechanisms regulating CRH-R2 functional activity and determining its biological responses.