title: Agonist-Activated Bombyx Corazonin Receptor Is Internalized via an Arrestin-Dependent and Clathrin-Independent Pathway.
abstract: Agonist-induced internalization plays a key role in the tight regulation of the extent and duration of G protein-coupled receptor signaling. Previously, we have shown that the Bombyx corazonin receptor (BmCrzR) activates both Gαq- and Gαs-dependent signaling cascades. However, the molecular mechanisms involved in the regulation of the internalization and desensitization of BmCrzR remain to be elucidated. Here, vectors for expressing BmCrzR fused with enhanced green fluorescent protein (EGFP) at the C-terminal end were used to further characterize BmCrzR internalization. We found that the BmCrzR heterologously expressed in HEK-293 and BmN cells was rapidly internalized from the plasma membrane into the cytoplasm in a concentration- and time-dependent manner via a β-arrestin (Kurtz)-dependent and clathrin-independent pathway in response to agonist challenge. While most of the internalized receptors were recycled to the cell surface via early endosomes, some others were transported to lysosomes for degradation. Assays using RNA interference revealed that both GRK2 and GRK5 were essentially involved in the regulation of BmCrzR phosphorylation and internalization. Further investigations indicated that the identified cluster of Ser/Thr residues ((411)TSS(413)) was responsible for GRK-mediated phosphorylation and internalization. This is the first detailed investigation of the internalization and trafficking of Bombyx corazonin receptors.
authors.affiliation: National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University , Zhoushan, Zhejiang 316022, China.|Department of Economic Zoology, College of Animal Sciences, Zhejiang University , Hangzhou, Zhejiang 310058, China.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Economic Zoology, College of Animal Sciences, Zhejiang University , Hangzhou, Zhejiang 310058, China.
authors.forename: Zhangfei
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Department of Economic Zoology, College of Animal Sciences, Zhejiang University , Hangzhou, Zhejiang 310058, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Jiang
authors.affiliation: College of Life Sciences, Zhejiang University , Zijingang Campus, Hangzhou, Zhejiang, China.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Yang
authors.affiliation: Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Science, Wenzhou Medical University , Wenzhou, Zhejiang 325035, China.
authors.forename: Haishan
authors.initials: H
authors.lastname: Huang
authors.affiliation: College of Life Sciences, Zhejiang University , Zijingang Campus, Hangzhou, Zhejiang, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Jin
authors.affiliation: Department of Economic Zoology, College of Animal Sciences, Zhejiang University , Hangzhou, Zhejiang 310058, China.
authors.forename: Yajie
authors.initials: Y
authors.lastname: Chen
authors.forename: Liangen
authors.initials: L
authors.lastname: Shi
authors.forename: Naiming
authors.initials: N
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019393:Arrestin / Q000378:metabolism*; D012831:Bombyx / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D034281:Dynamins / Q000378:metabolism; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism; D054774:G-Protein-Coupled Receptor Kinase 5 / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D021381:Protein Transport; D018013:Receptors, Neuropeptide / Q000819:agonists* / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D019393:Arrestin; D002966:Clathrin; D018013:Receptors, Neuropeptide; C468120:corazonin receptor; C496567:GRK2 protein, human; D054769:G-Protein-Coupled Receptor Kinase 2; D054774:G-Protein-Coupled Receptor Kinase 5; C517898:GRK5 protein, human; D034281:Dynamins