Cloning and expression of the cDNA encoding the FXPRL family of peptides and a functional analysis of their effect on breaking pupal diapause in Helicoverpa armigera.
Cloning and expression of the cDNA encoding the FXPRL family of peptides and a functional analysis of their effect on breaking pupal diapause in Helicoverpa armigera.
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DOI：
10.1016/j.jinsphys.2003.09.006
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发表时间：
2004
影响因子：
2.2
通讯作者：
Tian-Yi Zhang;Jiu-song Sun;Liu-Bin Zhang;Jin-Liang Shen;Wei-Hua Xu
中科院分区：
文献类型：
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作者：
Tian-Yi Zhang;Jiu-song Sun;Liu-Bin Zhang;Jin-Liang Shen;Wei-Hua Xu
滞育激素 (DH) 和信息素生物合成激活神经肽 (PBAN) 由食管下神经节 (SG) 中的单个 mRNA 编码，负责诱导家蚕胚胎滞育和鳞翅目昆虫性信息素生物合成。 PBAN cDNA 分析表明，DH 样肽存在于几种有蛹滞育的物种中。然而，DH 样肽的功能仍然未知。在本研究中，我们利用cDNA末端快速扩增方法克隆了棉铃虫中编码DH-PBAN的cDNA。核苷酸序列分析显示，该cDNA最长的开放阅读框编码194个氨基酸的前体蛋白，其中包含33个氨基酸的PBAN、24个氨基酸的DH样肽和其他三种神经肽，所有这些神经肽都具有共同的C端五肽基序FXPR/KL（X=G、T、S）。同源性检索表明，棉铃虫DH-like和PBAN与其他昆虫的DH-like和PBAN具有高度同源性。 Northern印迹分析表明，与来自蛹SG的Har-DH-PBAN cDNA大小相对应的单个信息RNA在非滞育蛹的SG中的表达显着高于滞育蛹。蛋白质印迹分析显示男性和女性的 SG 均表达 DH 样肽。当将 DH 样肽注射到非滞育幼虫和蛹中时，它不会诱导滞育，而是有效地破坏了棉铃虫的蛹滞育。 DH 终止蛹滞育的 ED50 为 20 pmol/蛹。来自 DH-PBAN 多蛋白前体的其他四种 FXPRLamide 神经肽具有滞育终止的交叉活性。因此，这些观察结果表明这些 FXPRL 家族肽在促进几种夜蛾物种的持续发育中具有潜在作用。该基因在噬菌体成虫和成虫中的高表达表明FXPRL家族肽可能具有多种生理功能。
Diapause hormone (DH) and pheromone biosynthesis activating neuropeptide (PBAN) are encoded by a single mRNA in the suboesophegeal ganglion (SG) and are responsible for induction of embryonic diapause in Bombyx mori and sex pheromone biosynthesis in lepidopteran insects. PBAN cDNA analyses revealed that the DH-like peptide is present in several species that have a pupal diapause. However, the function of the DH-like peptide remains unknown. In the present study, we cloned the cDNA encoding DH-PBAN in Helicoverpa armigera utilizing the rapid amplification of the cDNA ends method. The nucleotide se quence analysis revealed that the longest open reading frame of this cDNA encodes a 194-amino acid precursor protein that con tains a 33-aa PBAN, a 24-aa DH-like peptide, and three other neuropeptides, all of which have a common C-terminal pentapeptide motif FXPR/KL (X=G, T, S). A homology search showed that H. armigera DH-like and PBAN are highly homologous to those from other insects. Northern blot analysis demonstrated a single message RNA corresponding to the size of Har-DH-PBAN cDNA from pupal SG with significantly higher expression in the SG of nondiapause pupae than diapausing pupae. Western blot analysis showed DH-like peptide expression from SG of both males and females. When DH-like peptide was injected into nondiapause larvae and pupae, it did not induce diapause, but rather efficiently broke pupal diapause in H. armigera. The ED50of DH to terminate pupal diapause is 20 pmol/pupae. The other four FXPRLamide neuropeptides from the DH-PBAN polyprotein precursor have cross activity for diapause termination. These observations therefore suggest a potential role for these FXPRL family peptides in promoting continuous development in several noctuid species. The high expression of this gene in pharate adults and adults indicates that the FXPRL family peptides may have multiple physiological functions.