管纹艳虎天牛UDP-葡萄糖基转移酶基因的鉴定及表达特征分析 *

doi:10.7679/j.issn.2095-1353.2020.092

摘要/Abstract

摘要：

【目的】 昆虫UDP-葡萄糖基转移酶（UGT）在其内源性和外源性有毒化合物的解毒代谢过程中起着重要作用,但在管纹艳虎天牛Rhaphuma horsfieldi中UGT基因的研究尚属空白。【方法】 采用转录组学、生物信息学、进化和表达谱分析、蛋白同源建模等技术研究了管纹艳虎天牛的UGT基因家族。【结果】 从管纹艳虎天牛转录组中一共鉴定到36个RhorUGTs基因,其中17个具有全长序列。鞘翅目不同种间UGT基因数量的比较表明,管纹艳虎天牛具有中等数量的UGT基因。进化分析结果表明,RhorUGTs分布在10个亚家族中,其中UGT352亚家族为天牛科昆虫所特有,且该亚家族中RhorUGT2/7/10/16/18/27可能通过基因的复制产生。表达谱结果表明,大部分RhorUGTs基因在检测的所有组织中均有表达,部分基因呈现触角或跗节特异或高表达的特点,暗示它们具有嗅觉或触觉等功能。三级结构分析发现,RhorUGT17的α3、α4、β4和β5主要参与UDP-葡萄糖的结合。【结论】 本研究明确了管纹艳虎天牛UGT基因的数量、序列特征、进化关系及组织表达特征,研究结果为该种天牛解毒代谢机制的阐明奠定基础。

关键词: 管纹艳虎天牛, UDP-葡萄糖基转移酶, 进化分析, 表达谱, 同源建模

Abstract:

[Objectives] UDP-glucosyltransferase (UGT) in insects is of particular importance for the detoxification of endogenous and exogenous toxic compounds. However, no information about RhorUGT genes of Rhaphuma horsfieldi is available. [Methods] In this study, we characterized the UGT gene family from R. horsfieldi using transcriptomics, bioinformatics, phylogenic and expression profiling analyses, and homology modeling approaches. [Results] Based on the transcriptomic data, totally 36 RhorUGT genes were identified from this beetle, 17 of which were full-length sequences. A comparative study on UGT gene numbers among different coleopteran species revealed that R. horsfieldi harbored a medium UGT number. Phylogenetic analysis indicated that RhorUGTs were clustered into ten sub-families, with a cerambycid-specific one of UGT352. Notably, these members formed by RhorUGT2/7/10/16/18/27 from UGT352 were possibly derived from gene duplications. Expression profiles showed that most of RhorUGTs were detected in tested tissues and some were specifically or highly expressed in antennae or tarsi, suggesting their diverse functions including olfaction and touch. Results by three-dimensional structures showed that α3, α4, β4 and β5 of RhorUGT17 were mainly involved in the binding of UDP-glucose. [Conclusion] This study has identified the numbers, sequence characteristics, phylogenic relationships and tissue expression characteristics of R. horsfieldi UGTs, and thus provides reference data for addressing the detoxification mechanisms of this beetle.

Key words: Rhaphuma horsfieldi, UDP-glycosyltransferase, phylogenetic analysis, expression profile, homology modeling

王政全, 尹宁娜, 赵宁, 刘乃勇. 管纹艳虎天牛UDP-葡萄糖基转移酶基因的鉴定及表达特征分析 ^*[J]. 应用昆虫学报, 2020, 57(4): 898-910.

Zheng-Quan WANG, Ning-Na YIN, Ning ZHAO, Nai-Yong LIU. Identification and expression characterization of UDP-glucosyltransferase genes in Rhaphuma horsfieldi[J]. Chinese Journal of Applied Entomology, 2020, 57(4): 898-910.

图/表 6

Information for UGT genes in Rhaphuma horsfieldi

Comparison of UGT gene numbers of different sub-families in Coleoptera

图1 管纹艳虎天牛UGT的氨基酸序列比对三角形表示起催化作用的组氨酸（H）和天冬氨酸（D）;1、2和3分别表示与核苷酸、含磷化合物和糖苷作用的氨基酸。

Fig. 1 Multiple alignment of UGT amino acid sequences in Rhaphuma horsfieldi Triangle represents key catalytic residues (H, histidine and D, aspartic acid). 1, 2 and 3 represent nucleotide, phosphate and glucoside interacting residues, respectively.

图2 鞘翅目昆虫UGT的进化树

Fig. 2 Phylogenetic tree of UGTs in Coleoptera

图3 管纹艳虎天牛RhorUGTs基因在成虫不同组织的表达谱 FA：成虫触角;FTa：雌虫跗节;FB：不含触角和跗节的身体;MA：雄虫触角;MTa：雄虫跗节;MB：不含触角和跗节的身体。

Fig. 3 Expression pattern of RhorUGT genes in different tissues of Rhaphuma horsfieldi adults FA: Female antennae; FTa: Female tarsi; FB: Female bodies without antennae and tarsi; MA: Male antennae; MTa: Male tarsi; MB: Male bodies without antennae and tarsi.

图4 管纹艳虎天牛RhorUGT17的三级结构 A. 管纹艳虎天牛RhorUGT17与人HsapUGT2B7的氨基酸序列比对;B. 管纹艳虎天牛RhorUGT17的三级结构（左边）及其与HsapUGT2B7的三级结构叠加（右边）。两个蛋白的关键氨基酸（S、W、Q、T、H、E、D和H/Q）和UDP-葡萄糖结合腔在结构中被标示;供体结合区域（DBR1和DBR2）用黄色标示;Nt、Ct、α1-7和β1-6：分别表示氨基酸端、羧基端、α螺旋1至7和β折叠1至6。

Fig. 4 Three-dimensional (3D) structure of Rhaphuma horsfieldi UGT17 A. Alignment of RhorUGT17 and HsapUGT2B7 amino acid sequences; B. 3D model of RhorUGT17 (left) and superimposition of RhorUGT17 and HsapUGT2B7 structures (right). Key residues (S, W, Q, T, H, E, D and H/Q) and UDP-glucose binding cavities of RhorUGT17 and HsapUGT2B7 are labeled on the structures, respectively. Donor binding domains (DBR1 and DBR2) are highlighted in yellow. Nt, Ct, α1-7 and β1-6 represent N-terminus, C-terminus, seven α-helixes and six β-sheets, respectively.

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