Cloning and characterization of ribosomal protein S29, a deltamethrin resistance associated gene from Culex pipiens pallens

Sun, Haibo; Sun, Linchun; He, Ji; Shen, Bo; Yu, Jing; Chen, Chen; Yang, Mingxia; Sun, Yan; Zhang, Donghui; Ma, Lei; Zhu, Changliang

doi:10.1007/s00436-011-2443-z

Cited by 19 publications

(14 citation statements)

References 43 publications

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“…Ribosomal protein RPS29 was found to be associated with DM resistance in our previous study [14]. To investigate the mechanism, TAP was used to identify RPS29-interacting paterners.…”

Section: Discussionmentioning

confidence: 95%

“…Studies indicated that human RPS29 has extra-ribosomal functions as an apoptosis inducer and RNase [15], [16], but despite this, insect RPS29 remains poorly characterized. In our previous study, it had been identified that RPS29 is involved in DM resistance of mosquito [14].…”

Section: Introductionmentioning

confidence: 99%

“…One of these identified genes was ribosomal protein s29 ( rps29 ), which shares over 98% sequence identity with homologs in five other mosquito species, and 77% with the human homolog. The putative amino acid sequence of RPS29 is identical to that of Aedes albopictus , Culex quinquefasciatus and Aedes aegypti [14]. Ribosomal proteins are involved in protein synthesis in all living cells and are highly conserved from yeasts to mammals.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2014

PLoS ONE

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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.

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“…Ribosomal protein RPS29 was found to be associated with DM resistance in our previous study [14]. To investigate the mechanism, TAP was used to identify RPS29-interacting paterners.…”

Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

et al. 2014

PLoS ONE

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“…Cells were incubated with the transfection mixture for 4 h, and the transfection medium was replaced with 2 mL normal medium. After additional 24 h incubation, wells were then treated with various concentrations of deltamethrin (final concentrations, 0, 10 0.5 , 10 1.0 , 10 1.5 , and 10 2.0 μg/mL; Sun et al 2011). After another 48 h, 10 μL of CCK-8 solution were added to each well, and the plates were detected at 450 nm using a microplate reader after additional 4 h incubation.…”

Section: Methodsmentioning

confidence: 99%

Ribose-phosphate pyrophosphokinase 1 (PRPS1) associated with deltamethrin resistance in Culex pipiens pallens

et al. 2012

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Ribose-phosphate pyrophosphokinase 1 (PRPS1) was identified and isolated as a differentially expressed gene between deltamethrin-susceptible (DS) and deltamethrin-resistant (DR) Culex pipiens pallens and Aedes albopictus C6/36 cell line through microarray and 2D-Gel. An open reading frame of PRPS1 cloned from C. pipiens pallens has 1,011 bp and encodes for a 336 amino acids protein which shares high homology with Culex quinquefasciatus. Real-time polymerase chain reaction was used to determine the transcript expression level of PRPS1 in DS and DR strains. The expression levels of PRPS1 were higher in DR laboratory strains and natural population JXZ-DR, JXZ-LDR. PRPS1 was also detected and expressed at all developmental stages of C. pipiens pallens and increased expression level in DR3 strain than DS strain in the third and fourth instar larvae, female and male stages. In addition, to further investigate the role of PRPS1 in deltamethrin resistance, PRPS1 was transiently expressed in A. albopictus C6/36 cells and detected by western blotting. Cells transfected with PRPS1 had an increased resistance to deltamethrin compared with control cells. These results suggested that the increased expression level of PRPS1 may play roles in the regulation of deltamethrin resistance.

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“…C. pipiens mücadelesinde özellikle organofosfat temelli kimyasallar ve böcek büyüme inhibitörleri yaygın bir şekilde kullanılmaktadır. Fakat bu kimyasalların çevre ve insan sağlığı üzerindeki yan etkisi ve zararlıların uzun vade de bu kimyasallara karşı direnç geliştirmesi gibi kaygıları da beraberinde getirmektedir (Sun et al, 2011;Azizoğlu ve ark., 2012;Azizoğlu, 2014;Benelli et al, 2014;Lees et al, 2014). Bu kaygılardan dolayı doğal ortamla daha uyumlu, insan ve çevre sağlığı açısından daha güvenli mücadele tekniklerinin geliştirilmesi her geçen gün daha da önem kazanmaktadır.…”

Section: Introductionunclassified

Investigation of Potential Usability of Native Bacillus thuringiensis Isolates in Biological Control of Disease Vector, Culex pipiens (Diptera:Culicidae)

Azizoğlu¹

2017

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ÖZET: Culex pipiens (Diptera: Culicidae) Batı Nil Virüs'ünün başlıca vektörüdür. Kimyasal pestisit kullanımının ve C. pipiens kaynaklı hastalıkların azaltılması için entomopatojen bakteri Bacillus thuringiensis (Bt) ürünleri yıllardır etkili bir şekilde kullanılmaktadır. Bu çalışmada, 14 farklı yerel Bt izolatının C. pipiens larvaları üzerindeki biyolojik mücadele potansiyeli araştırılmıştır. Dipteran spesifik cry gen taşıyan 14 farklı Bt izolatının spor-kristal protein karşımı (500 µg ml -1 ) C. pipens'in son dönem larvalarına uygulanmıştır. Uygulanan izolatlar arasında Bt SY50.4'ün spor-kristal protein karşımı % 80 larval ölüme sebep olmuştur. Bt SY56.3 izolatı, Bt SY50.4 kadar etkili olmasa da kontrolden daha fazla ölüm oranına sebep olmuştur. Diğer izolatlar C. pipens larvaları üzerinde önemli bir etki göstermemiştir. Bt SY50.4 izolatının C. pipiens üzerindeki etkinliği göz önüne alındığında biyolojik mücadele açısından gelecek vaat ettiği belirlenmiştir. Uygulama sonrası yaşayan larvaların pupalaşma ve erginleşme süreçleri kontrol grubu ile karşılaştırıldığında aralarında önemli bir fark olmadığı belirlenmiştir. Anahtar kelimeler: Bacillus thuringiensis, biyolojik mücadele, Culex pipiens, hayat döngüsüABSTRACT: Culex pipiens (Diptera: Culicidae) is primary vector of West Nile virus. Products of entomopathogen bacterium Bacillus thuringiensis (Bt) has effectively been used for decades for decreasing the chemical input into the environment and C. pipiens mediated diseases. In the current study, biocontrol potential of 14 different native Bt isolates was investigated on C. pipiens larvae. Last instar larvae of C. pipens were subjected to spore-crystal protein mixture (500 µg ml -1 ) of 14 different Bt isolates carrying dipteran active cry genes. Of all the tested isolates, the products of Bt SY50.4 caused 80% mortality on the larvae. Although Bt SY56.3 showed lower insecticidal effect than that of Bt SY50.4, the activity of this isolate was higher than control. The other tested isolates didn't show significant mortality on larvae of the C. pipiens. Bt SY50.4 was determined as promising an isolate based on the effectiveness on the C. pipiens larvae. Pupation and adult emergence periods of remaining larvae after treatment were not significantly different when compared with the control.

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Cloning and characterization of ribosomal protein S29, a deltamethrin resistance associated gene from Culex pipiens pallens

Cited by 19 publications

References 43 publications

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

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

Ribose-phosphate pyrophosphokinase 1 (PRPS1) associated with deltamethrin resistance in Culex pipiens pallens

Investigation of Potential Usability of Native Bacillus thuringiensis Isolates in Biological Control of Disease Vector, Culex pipiens (Diptera:Culicidae)

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