Department of Biology, National Changhua University of Education, Changhua 50058, Taiwan.
Gen Comp Endocrinol. 2013 Jun 1;186:80-4. doi: 10.1016/j.ygcen.2013.03.010. Epub 2013 Mar 18.
It was demonstrated in a previous study (Wu et al., 2012b) that crustacean hyperglycemic hormone (CHH) gene is expressed in the hemocyte of Procambarus clarkii. In the present study, 2 additional cDNAs (CHH2-L and tCHH2) from the hemocyte and a CHH gene (CHH2) from the abdominal muscle of the same species were cloned. Analyses of the cDNA and genomic sequences suggested that, similar to other previously reported CHH genes, 2 precursor transcripts (CHH2 and CHH2-L) would be derived from CHH2 gene through a process of RNA alternative splicing, and CHH2 and CHH2-L each encode a precursor containing a signal peptide, a CHH precursor-related peptide, and a mature peptide. Further, tCHH2 sequence consists of exon I, exon II, and a truncated segment of intron II of CHH2 gene, followed by a previously unknown 3'sequence. It is suggested that, because the truncation disrupts the highly conserved RNA splice acceptor site, the truncated segment is retained within tCHH2, resulting in encoding a precursor containing the typical precursor components except the mature peptide is truncated with only 40 residues. In addition, unlike 2 other previously identified transcripts (referred to as CHH1 and CHH1-L), CHH2-L, CHH2, tCHH2 contain in the 3'-UTRs 3-5 AU-rich elements (AREs). The data showed that multiple CHH genes are expressed in crayfish hemocytes. Novel sequence characteristics of the transcripts result in an RNA splicing pattern that yields a transcript (tCHH2) encoding a precursor with an atypical truncated mature peptide and possibly leads to a different expression dynamics of the precursors encoded by the ARE-containing transcripts.
先前的研究表明,甲壳动物血糖激素(CHH)基因在克氏原螯虾的血细胞中表达。在本研究中,从相同物种的血细胞中克隆了另外 2 个 cDNA(CHH2-L 和 tCHH2)和 1 个来自腹肌肉的 CHH 基因(CHH2)。对 cDNA 和基因组序列的分析表明,与其他先前报道的 CHH 基因类似,2 个前体转录物(CHH2 和 CHH2-L)将通过 RNA 选择性剪接过程从 CHH2 基因衍生而来,CHH2 和 CHH2-L 各自编码一个含有信号肽、CHH 前体相关肽和成熟肽的前体。此外,tCHH2 序列由 CHH2 基因的外显子 I、外显子 II 和内含子 II 的截断片段组成,后面是一个先前未知的 3'序列。这表明,由于截断破坏了高度保守的 RNA 剪接受体位点,截断片段保留在 tCHH2 中,导致编码一个含有典型前体成分的前体,除了成熟肽被截断仅有 40 个残基。此外,与另外 2 个先前鉴定的转录物(称为 CHH1 和 CHH1-L)不同,CHH2-L、CHH2、tCHH2 在 3'-UTR 中含有 3-5 个 AU 丰富元件(AREs)。数据表明,多个 CHH 基因在螯虾血细胞中表达。这些转录物的新型序列特征导致 RNA 剪接模式产生一个编码具有非典型截断成熟肽的前体的转录物(tCHH2),并可能导致含有 ARE 的转录物编码的前体的表达动力学不同。