Cloning and Characterization of 5′-Upstream Sequence of the M32 Gene for a Mouse Homologue of Drosophila Heterochromatin Protein 1 (HP1)

M. Sato, K. Miyado, M. Kimura
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引用次数: 7

Abstract

M32 [also termed chromatin modifier protein 2 (MOD2)] is a nuclear protein consisting of the condensed chromatin structure (heterochromatin) and considered one of the mammalian homologues of heterochromatin protein 1 (HP1), first isolated as one of the components of heterochromatin in Drosophila. This report presents the isolation and characterization of the 5′-upstream region of the mouse M32 gene containing a promoter region and 5′-untranslated region (5′-UTR) exon. The 5′-upstream region (approximately 0.27 kb starting from the 5′ end of the 5′-UTR exon) of the M32 gene contained neither a TATA box nor a CCAAT box, but possessed potential binding sites for transcription factors such as Spl, H4TF-1, PEA2, PEA3, GSG element and Egr-1, and was highly G/C-rich. The promoter activity of this 5′-upstream region was demonstrated by transfecting its fusion-construct with the E. coli β-galactosidase gene into the F9 mouse teratocarcinoma cell line. The 5′ ends of the mRNA were mapped to at least two positions in the 5′-upstream region. Interestingly, the 5′-upstream region exhibited a high degree of similarity to a portion of heterogeneous nuclear ribonucleo-protein (hnRNP) A2/B1 gene, which is thought to play a role in RNA processing, located in the reverse orientation to the M32 gene, and also to several known ESTs and cDNAs. These findings suggest that the 5′-upstream region of the M32 gene consists of a multiple regulatory complex which probably plays important roles in nuclear function such as chromatin organization and RNA processing.
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果蝇异染色质蛋白1 (HP1)同源物M32基因5′上游序列的克隆与鉴定
M32[也称为染色质修饰蛋白2 (chromatin modifier protein 2, MOD2)]是一种由凝聚染色质结构(heterochromatin)组成的核蛋白,被认为是异染色质蛋白1 (heterochromatin protein 1, HP1)的哺乳动物同源物之一,最早在果蝇中作为异染色质成分之一被分离出来。本文报道了小鼠M32基因的5 '上游区域的分离和表征,该区域包含一个启动子区域和5 ' -未翻译区(5 ' -UTR)外显子。M32基因的上游5′区(从5′-UTR外显子5′端开始约0.27 kb)既不含TATA box,也不含CCAAT box,但具有Spl、H4TF-1、PEA2、PEA3、GSG element和Egr-1等转录因子的潜在结合位点,且G/ c含量高。通过将其与大肠杆菌β-半乳糖苷酶基因的融合构建体转染到F9小鼠畸胎癌细胞系中,证实了该5 '上游区域的启动子活性。mRNA的5 '端被定位到5 '上游区域的至少两个位置。有趣的是,5 ' -上游区域与异质核糖核蛋白(hnRNP) A2/B1基因的一部分高度相似,该基因被认为在RNA加工中起作用,位于与M32基因相反的方向,也与几种已知的est和cdna相似。这些发现表明,M32基因的5 '上游区域由一个多调控复合体组成,该复合体可能在染色质组织和RNA加工等核功能中起重要作用。
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