Kwon Deok Ho, Kim Ju Hyeon, Kim Young Ho, Yoon Kyong Sup, Clark J Marshall, Lee Si Hyeock
Research Institute for Agriculture and Life Sciences, Seoul National University, 151-921, Republic of Korea.
Department of Agricultural Biotechnology, Seoul National University, 151-921, Republic of Korea.
Pestic Biochem Physiol. 2014 Jun;112:13-8. doi: 10.1016/j.pestbp.2014.05.006. Epub 2014 Jun 7.
Enhanced malathion carboxylesterase (MCE) activity was previously reported to be involved in malathion resistance in the head louse Pediculus humanus capitis (Gao et al., 2006 [8]). To identify MCE, the transcriptional profiles of all five esterases that had been annotated to be catalytically active were determined and compared between the malathion-resistant (BR-HL) and malathion-susceptible (KR-HL) strains of head lice. An esterase gene, designated HLCbE3, exhibited approximately 5.4-fold higher transcription levels, whereas remaining four esterases did not exhibit a significant increase in their transcription in BR-HL, indicating that HLCbE3 may be the putative MCE. Comparison of the entire cDNA sequences of HLCbE3 revealed no sequence differences between the BR-HL and KR-HL strains and suggested that no single nucleotide polymorphism is associated with enhanced MCE activity. Two copies of the HLCbE3 gene were observed in BR-HL, implying that the over-transcription of HLCbE3 is due to the combination of a gene duplication and up-regulated transcription. Knockdown of HLCbE3 expression by RNA interference in the BR-HL strain led to increases in malathion susceptibility, confirming the identity of HLCbE3 as a MCE responsible for malathion resistance in the head louse. Phylogenetic analysis suggested that HLCbE3 is a typical dietary esterase and belongs to a clade containing various MCEs involved in malathion resistance.
先前有报道称,增强的马拉硫磷羧酸酯酶(MCE)活性与头虱(人头虱)对马拉硫磷的抗性有关(Gao等人,2006年[8])。为了鉴定MCE,测定并比较了头虱抗马拉硫磷(BR-HL)品系和敏感马拉硫磷(KR-HL)品系中所有五个已注释具有催化活性的酯酶的转录谱。一个名为HLCbE3的酯酶基因在BR-HL中的转录水平高出约5.4倍,而其余四个酯酶在BR-HL中的转录没有显著增加,这表明HLCbE3可能是推定的MCE。对HLCbE3的完整cDNA序列进行比较,发现BR-HL和KR-HL品系之间没有序列差异,这表明没有单核苷酸多态性与增强的MCE活性相关。在BR-HL中观察到两个拷贝的HLCbE3基因,这意味着HLCbE3的过度转录是由于基因复制和转录上调共同作用的结果。通过RNA干扰在BR-HL品系中敲低HLCbE3的表达导致对马拉硫磷的敏感性增加,证实了HLCbE3是导致头虱对马拉硫磷产生抗性的MCE。系统发育分析表明,HLCbE3是一种典型的膳食酯酶,属于一个包含各种与马拉硫磷抗性相关的MCE的进化枝。