DaTrypsin, a novel clip-domain serine proteinase gene up-regulated during winter and summer diapauses of the onion maggot, Delia antiqua
DaTrypsin, a novel clip-domain serine proteinase gene up-regulated during winter and summer diapauses of the onion maggot, Delia antiqua
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DOI：
10.1016/j.gene.2004.12.026
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发表时间：
2005-02-28
期刊：
影响因子：
3.5
通讯作者：
Ishikawa, Y
中科院分区：
文献类型：
--
作者：
Chen, B;Kayukawa, T;Ishikawa, Y
滞育使昆虫和其他节肢动物做好在恶劣环境中生存的准备。为了探索冬季（WD）和夏季滞育（SD）的分子基础，我们在洋葱蛆（Delia antiqua）中筛选了滞育特异性基因，该蛆在夏季和冬季都以蛹的形式滞育。通过差异显示鉴定滞育诱导的转录物 DaTrypsin，并通过 Northern 印迹、定量实时 PCR 和序列分析进行检查。全长 cDNA，长 1379 bp，编码 384 个氨基酸。分子量为 43,005 Da。该蛋白质含有 20 个氨基酸。分泌肽，随后是氨基末端夹结构域和羧基末端丝氨酸蛋白酶结构域。以 Ser、His 和 Asp 作为催化残基，以 Asp、Gly 和 Ser 作为特异性决定簇，DaTrypsin 有望成为一种胰蛋白酶样酶。 DaTrypsin 转录在 SD 和 WD 蛹中均上调，WD 期间的 mRNA 水平高于 SD。热休克进一步提高了SD和WD蛹中的基因转录，而冷休克降低了SD蛹中DaTrypsin的表达，并且对WD蛹没有显着影响。在 SD 蛹中，DaTrypsin 转录物在滞育期间和温度冲击后逐渐增加，而在 WD 蛹中，DaTrypsin mRNA 水平在滞育开始时和温度冲击后立即很高，然后随着时间的推移逐渐降低。 DaTrypsin 代表第一个在滞育期间表达的丝氨酸蛋白酶基因，也是第一个在 SD 和 WD 中上调的基因。它可能参与宿主的免疫防御和/或维持滞育蛹的发育状态。 (c) 2004 Elsevier B.V. 保留所有权利。
Diapause prepares insects and other arthropods to survive in harsh environments. To explore the molecular basis of winter (WD) and summer diapauses (SD), we screened for diapause-specific genes in the onion maggot, Delia antiqua, that diapauses as a pupa in both summer and winter. A diapause-induced transcript, DaTrypsin, was identified through differential display, and examined by Northern blot, quantitative real-time PCR and sequence analyses. The full-length cDNA, 1379 bp long, encodes 384 a.a. with a molecular mass of 43,005 Da. The protein contains a 20-a.a. secretion peptide, followed by an amino-terminal clip domain and a carboxyl-terminal serine proteinase domain. With Ser, His and Asp as catalytic residues and Asp, Gly and Ser as specificity determinants, DaTrypsin is anticipated to be a trypsin-like enzyme. DaTrypsin transcription is up-regulated in both SD and WD pupae with higher mRNA levels during WD than SD. Heat shock further elevated gene transcription in both SD and WD pupae, whereas cold shock reduced DaTrypsin expression in SD pupae and had no significant effect on WD pupae. In SD pupae, DaTrypsin transcripts gradually build up during diapause, and after temperature shocks, whereas in WD pupae DaTrypsin mRNA levels are high at the beginning of diapause and immediately after a temperature shock and then gradually decrease with time. DaTrypsin represents the first serine proteinase gene expressed during diapause as well as the first gene upregulated in both SD and WD. It may participate in the host's immune defense and/or maintain the developmental status in the diapausing pupae. (c) 2004 Elsevier B.V. All rights reserved.