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红带锥蝽中的ACBP基因家族:RpACBP-1的表达、特性及功能

The ACBP gene family in Rhodnius prolixus: Expression, characterization and function of RpACBP-1.

作者信息

Majerowicz David, Hannibal-Bach Hans K, Castro Rodolfo S C, Bozaquel-Morais Bruno L, Alves-Bezerra Michele, Grillo Luciano A M, Masuda Claudio A, Færgeman Nils J, Knudsen Jens, Gondim Katia C

机构信息

Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Institut for Biokemi og Molekylær Biologi, Syddansk Universitet, Odense, Denmark.

Institut for Biokemi og Molekylær Biologi, Syddansk Universitet, Odense, Denmark.

出版信息

Insect Biochem Mol Biol. 2016 May;72:41-52. doi: 10.1016/j.ibmb.2016.03.002. Epub 2016 Mar 19.

Abstract

The acyl-CoA-binding proteins (ACBP) constitute a family of conserved proteins that bind acyl-CoA with high affinity and protect it from hydrolysis. Thus, ACBPs may have essential roles in basal cellular lipid metabolism. The genome of the insect Rhodnius prolixus encodes five ACBP genes similar to those described for other insect species. The qPCR analysis revealed that these genes have characteristic expression profiles in insect organs, suggesting that they have specific roles in insect physiology. Recombinant RpACBP-1 was able to bind acyl-CoA in an in vitro gel-shift assay. Moreover, heterologous RpACBP-1 expression in acb1Δ mutant yeast rescued the multi-lobed vacuole phenotype, indicating that RpACBP-1 acts as a bona fide acyl-CoA-binding protein. RpACBP-1 knockdown using RNAi caused triacylglycerol accumulation in the insect posterior midgut and a reduction in the number of deposited eggs. The amount of stored triacylglycerol was reduced in flight muscle, and the incorporation of fatty acids in cholesteryl esters was increased in the fat body. These results showed that RpACBP-1 participates in several lipid metabolism steps in R. prolixus.

摘要

酰基辅酶A结合蛋白(ACBP)构成了一个保守蛋白家族,它们能以高亲和力结合酰基辅酶A并保护其不被水解。因此,ACBP可能在基础细胞脂质代谢中发挥重要作用。昆虫长红猎蝽的基因组编码了五个ACBP基因,与其他昆虫物种中描述的基因相似。定量PCR分析表明,这些基因在昆虫器官中具有特征性的表达谱,这表明它们在昆虫生理学中具有特定作用。在体外凝胶迁移试验中,重组RpACBP-1能够结合酰基辅酶A。此外,在acb1Δ突变酵母中异源表达RpACBP-1挽救了多叶液泡表型,表明RpACBP-1作为一种真正的酰基辅酶A结合蛋白发挥作用。使用RNA干扰敲低RpACBP-1会导致昆虫中肠后部甘油三酯积累以及产卵数量减少。飞行肌中储存的甘油三酯量减少,脂肪体中脂肪酸掺入胆固醇酯的量增加。这些结果表明,RpACBP-1参与了长红猎蝽的多个脂质代谢步骤。

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