Identification of a melanocyte-type promoter of the microphthalmia-associated transcription factor gene
Identification of a melanocyte-type promoter of the microphthalmia-associated transcription factor gene
复制标题
DOI：
10.1006/bbrc.1996.0298
复制
发表时间：
1996-02-27
影响因子：
3.1
通讯作者：
Shibahara, S
中科院分区：
文献类型：
--
作者：
Fuse, N;Yasumoto, K;Shibahara, S
小眼球相关转录因子(MITF)是小鼠小眼症基因产物的人类同源物，调节黑素细胞特异性酪氨酸酶基因的转录，该基因编码黑色素生物合成中的一种关键酶。在本研究中，我们克隆并鉴定了人类基因组DNA片段，该片段包含MITF基因的一个黑素细胞型外显子及其5‘-侧翼区。通过使用黑色素瘤RNA的引物延伸和S1核酸酶作图分析，确定了一个主要的转录起始点。随后，构建了含有萤火虫荧光素酶基因上游5‘侧翼区的融合基因，并将其导入不表达MITF mRNA的黑色素瘤细胞或HeLa细胞。瞬时表达分析表明，2.3kb的5‘侧翼区能够使荧光素酶基因在色素细胞中优先表达。这些结果证实了MITF基因含有黑素细胞特异性启动子。(C)1996年学术出版社。
Microphthalmia-associated transcription factor (MITF), the human homolog of the mouse microphthalmia gene product, regulates melanocyte-specific transcription of the tyrosinase gene that codes for an essential enzyme in melanin biosynthesis. In this study, we have cloned and characterized the human genomic DNA segment containing a melanocyte-type exon and its 5'-flanking region of the MITF gene. A major transcriptional initiation site was assigned by primer extension and S1 nuclease mapping analysts using melanoma RNA. Subsequently, the fusion genes, containing the identified 5'-flanking region upstream from the firefly luciferase gene, were constructed and were introduced into pigmented melanoma cells or HeLa cells which do not express MITF mRNA. Transient expression assays show that the 5'-flanking region of 2.3 kb is able to confer preferential expression of a luciferase gene in pigment cells. These results establish that the MITF gene contains a melanocyte-specific promoter. (C) 1996 Academic Press, Inc.