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鸡羟基吲哚-O-甲基转移酶基因的结构分析

Structural analysis of the chicken hydroxyindole-O-methyltransferase gene.

作者信息

Grechez-Cassiau A, Bernard M, Ladjali K, Rodriguez I R, Voisin P

机构信息

Laboratory of Cellular Neurobiology, UMR CNRS 6558, UFR Sciences, Poitiers, France.

出版信息

Eur J Biochem. 1998 Nov 15;258(1):44-52. doi: 10.1046/j.1432-1327.1998.2580044.x.

Abstract

Hydroxyindole-O-methyltransferase (HIOMT) catalyzes the final step of melatonin synthesis, a neurohormone involved in photoperiodism and produced specifically in the pineal gland and in the retina. In the chicken, HIOMT gene transcription appears to be controlled by a circadian oscillator located in the pineal gland. We have characterized the chicken HIOMT gene over 17 kb, including 2.9 kb of 5'-flanking sequence. The major transcript (1.6 kb) is composed of eight exons distributed over 7.5 kb of genomic DNA. A ninth alternative exon was identified 6 kb downstream of exon 8. It was found in minor transcripts in the pineal gland and in the retina. Sequence similarity between exons 8 and 9 suggests their origin by exon duplication. Due to early stop codons, inclusion of exon 9 truncates the open reading frame by up to 33%. A restriction fragment length polymorphism was detected for a BglII site in intron 8. Fluorescence hybridization localized the HIOMT gene on chicken chromosome 1q22. The 5'-flanking region contains GATTAA and TAATCC sequences that may be related to tissue-specific expression. An ATTTAAAT sequence at position -29 would play the role of a TATA box, as evidenced by electrophoretic mobility shift assay. Information obtained in this study open the way to further studies aimed at analyzing the circadian rhythm of transcription at promoter level.

摘要

羟基吲哚 - O - 甲基转移酶(HIOMT)催化褪黑素合成的最后一步,褪黑素是一种参与光周期调节的神经激素,专门在松果体和视网膜中产生。在鸡中,HIOMT基因转录似乎受位于松果体中的昼夜节律振荡器控制。我们已经对超过17 kb的鸡HIOMT基因进行了特征分析,包括2.9 kb的5'侧翼序列。主要转录本(1.6 kb)由分布在7.5 kb基因组DNA上的八个外显子组成。在第8外显子下游6 kb处鉴定出第九个可变外显子。它存在于松果体和视网膜的次要转录本中。第8和第9外显子之间的序列相似性表明它们起源于外显子重复。由于早期终止密码子,包含第9外显子会使开放阅读框截断多达33%。在第8内含子的一个BglII位点检测到限制性片段长度多态性。荧光杂交将HIOMT基因定位在鸡的1号染色体1q22上。5'侧翼区域包含可能与组织特异性表达相关的GATTAA和TAATCC序列。位置 - 29处的ATTTAAAT序列可能起到TATA盒的作用,电泳迁移率变动分析证明了这一点。本研究获得的信息为进一步研究在启动子水平分析转录的昼夜节律开辟了道路。

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