title: Activation of Bombyx neuropeptide G protein-coupled receptor A4 via a Gαi-dependent signaling pathway by direct interaction with neuropeptide F from silkworm, Bombyx mori.
abstract: Members of the mammalian neuropeptide Y (NPY) family serve as neurotransmitters and contribute to a diversity of physiological functions. Although neuropeptide F (NPF), the NPY-like orthologs from insects, have been identified, the NPF receptors and their signaling and physiological functions remain largely unknown. In this study, we established the stable and transient functional expression of a Bombyx orphan G protein-coupled receptor, BNGR-A4, in both mammalian HEK293 and insect SF21 cells. We identified Bombyx mori NPFs as specific endogenous ligands for the Bombyx Neuropeptide GPCR A4 (BNGR-A4) and, accordingly, named the receptor BomNPFR. Our results demonstrated that BomNPFR was activated by synthetic BomNPF1a and BomNPF1b at a high efficacy and by BomNPF2 at a low efficacy. This activation led to a decrease of forskolin or adipokinetic hormone peptide-stimulated adenylyl cyclase activity, an increase of intracellular Ca(2+), the activation of ERK1/2 signaling and receptor internalization. Moreover, a Rhodamine-labeled BomNPF1a peptide was found to bind specifically to BomNPFR. The results derived from quantitative RT-PCR analysis and dsRNA-mediated knockdown experiments demonstrated the possible role of BomNPFR in the regulation of food intake and growth. Our results provide the first in-depth information on BomNPFR-mediated signaling for the further elucidation of the BomNPF/BomNPFR system in the regulation of fundamental physiological processes.
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Deng
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Yang
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Xiaobai
authors.initials: X
authors.lastname: He
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liao
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Congxia
authors.initials: C
authors.lastname: Zheng
authors.affiliation: Zhejiang Provincial Key Laboratory for Model Organisms, School of Life Sciences, Wenzhou Medical College, Wenzhou, Zhejiang 325035, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China.
authors.forename: Chenggang
authors.initials: C
authors.lastname: Zhu
authors.affiliation: Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu 410127, China.
authors.forename: Guozheng
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Zhejiang Provincial Key Laboratory for Model Organisms, School of Life Sciences, Wenzhou Medical College, Wenzhou, Zhejiang 325035, China.
authors.forename: Jimin
authors.initials: J
authors.lastname: Gao
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang Campus, Zhejiang University, Hangzhou, Zhejiang 310058, China. Electronic address: znm2000@yahoo.com.
authors.forename: Naiming
authors.initials: N
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D012831:Bombyx / Q000378:metabolism*; D003001:Cloning, Molecular; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D018013:Receptors, Neuropeptide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D061987:Sf9 Cells; D015398:Signal Transduction
chemical_list: D019476:Insect Proteins; D018013:Receptors, Neuropeptide
keywords: Arrestin; GPCR; MAP kinases; Neuropeptide; Signaling