Ribosomal Protein S29 Regulates Metabolic Insecticide Resistance through Binding and Degradation of CYP6N3

Abstract

BackgroundMany 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.Methodology and Principal FindingsIn 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 and SignificanceThese 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.

Proteomic analysis of Bayvarol® resistance mechanisms in the honey bee parasite Varroa destructor

“…Deltamethrin resistant mosquitoes demonstrated a 23-fold increase in the expression of the L39 gene (Tan et al, 2007), but this was not the case in the Bayvarol ® resistant mites studied here. Yu et al (2014) demonstrated that another ribosomal protein s29 regulates metabolic resistance by binding to CYP6N3, so perhaps the unbound versions of certain ribosomal proteins were more abundant during proteomic analysis of the sensitive mites.…”

CYP4BN6 and CYP6BQ11 mediate insecticide susceptibility and their expression is regulated by Latrophilin in Tribolium castaneum

“…In particular, reinforced enzymatic detoxification is one of the most common methods of achieving insecticide resistance of insects . Enzymatic detoxification is identified as a metabolic resistance process that involves the biochemical transformation of insecticides and further increases the biodegradation of toxic insecticides . This generally occurs due to the continuously increased expression and/or induction and elevated activity of members of three detoxification enzyme families: cytochrome P450 proteins (CYPs), carboxylesterases (CarEs), and glutathione S‐transferases (GSTs) .…”

“…This generally occurs due to the continuously increased expression and/or induction and elevated activity of members of three detoxification enzyme families: cytochrome P450 proteins (CYPs), carboxylesterases (CarEs), and glutathione S‐transferases (GSTs) . It has been shown that CYPs are the primary enzyme family that is associated with resistance to chemical insecticides in insect species . To date, a total of 143 CYPs, which can be assigned to four clans (clan 4, clan 3, clan 2, and a mitochondrial clan), have been identified in T. castaneum .…”

“…The use of common chemical insecticides makes it possible to employ more effective management strategies for pest control. 1,2 However, excessive and continuous application of insecticides has resulted in the development of insecticide resistance in many insect species, 3 and this has become a principal obstacle to control the outbreak of disaster pests, some of which are resurgent. Insecticide resistance of insects usually operates through complex polygenic inheritance, the mechanisms of which are not well understood.…”

“…12 Enzymatic detoxification is identified as a metabolic resistance process that involves the biochemical transformation of insecticides and further increases the biodegradation of toxic insecticides. 2,11 This generally occurs due to the continuously increased expression and/or induction and elevated activity of members of three detoxification enzyme families: cytochrome P450 proteins (CYPs), carboxylesterases (CarEs), and glutathione S-transferases (GSTs). [13][14][15] It has been shown that CYPs are the primary enzyme family that is associated with resistance to chemical insecticides in insect species.…”