title: Ribosomal protein S29 regulates metabolic insecticide resistance through binding and degradation of CYP6N3.
abstract: BACKGROUND
  Many diseases are transmitted by mosquitoes, including malaria, dengue fever, yellow fever, filariasis, and West Nile fever. Chemical control plays a major role in managing mosquito-borne diseases. However, excessive and continuous application of insecticides has caused the development of insecticide resistance in many species including mosquito, and this has become the major obstacle to controlling mosquito-borne diseases. Insecticide resistance is the result of complex polygenic inheritance, and the mechanisms are not well understood. Ribosomal protein RPS29 was found to be associated with DM resistance in our previous study. In this study, we aim to further investigate the involvement of RPS29 in deltamethrin resistance.
  
  
  RESULTS
  In this study, tandem affinity purification was used to identify proteins that can interact with RPS29. Among the candidate proteins, CYP6N3, a member of the CYP450 superfamily, was identified, and binding to RPS29 was confirmed in vitro and in vivo by GST pull-down and immunofluorescence. CCK-8 assay was used to investigate the RPS29-CTP6N3 interaction in relation to DM resistance. CYP6N3 overexpression significantly enhanced DM resistance and insect cell viability, but this was reversed by RPS29 overexpression. Western blot was used to study the mechanism of interaction between RPS29 and CYP6N3. RPS29 increases CYP6N3 protein degradation through the proteasome.
  
  
  CONCLUSIONS
  These observations indicate that CYP6N3, a novel RPS29-interacting partner, could stimulate deltamethrin resistance in mosquito cells and RPS29 overexpression targeted CYP6N3 for proteosomal degradation, abrogating the CYP6N3-associated resistence to deltamethrin. Our findings provide a novel mechanism associated with CYP450s mediated DM resistance.
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Shengli
authors.initials: S
authors.lastname: Hu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Ma
authors.affiliation: Pediatric Research Center, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.
authors.forename: Linchun
authors.initials: L
authors.lastname: Sun
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Hu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Zou
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Zhentao
authors.initials: Z
authors.lastname: Lei
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Donghui
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Shen
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D009033:Culicidae; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D020868:Gene Silencing; D007305:Insecticide Resistance / Q000235:genetics; D007306:Insecticides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D059748:Proteolysis; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D002352:Carrier Proteins; D007306:Insecticides; D012269:Ribosomal Proteins; C079855:ribosomal protein S29; D003577:Cytochrome P-450 Enzyme System; D046988:Proteasome Endopeptidase Complex
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