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非洲爪蟾卵母细胞和胚胎中c-myc启动子活性的调控

Regulation of Xenopus c-myc promoter activity in oocytes and embryos.

作者信息

Modak S P, Principaud E, Spohr G

机构信息

Université de Genève, Département de Biologie Cellulaire, Geneva, Switzerland.

出版信息

Oncogene. 1993 Mar;8(3):645-54.

PMID:8437848
Abstract

We have studied the regulation of transcription of the Xenopus c-myc I gene in oocytes and embryos. Various 5' and internal deletions of a 1310-bp-long c-myc I promoter fragment have been ligated upstream of the chloramphenicol acetyl transferase (CAT) reporter gene and microinjected into oocytes and fertilized eggs. Activity was determined by CAT assay and primer extension. The c-myc promoter drives transcription very efficiently, and a truncated promoter -158/+46 essentially retains full activity. This region contains an overlapping E2F/SP1 site and two tandem Sp1 sites homologous to those found in the c-myc gene of mouse. Internal deletions show that both elements are equally active in oocytes in driving the expression of CAT. A germinal vesicle extract contains a DNA-binding activity specific for an Sp1 consensus sequence but not the E2F site. The data suggest that the high transcription level of the endogenous c-myc gene in Xenopus oocytes is mediated by Sp1 or a related transcription factor. In embryos a different mechanism emerges and the functional role of the Sp1 binding sites appears to be less important.

摘要

我们研究了非洲爪蟾c-myc I基因在卵母细胞和胚胎中转录的调控。将一个1310bp长的c-myc I启动子片段的各种5'端和内部缺失片段连接到氯霉素乙酰转移酶(CAT)报告基因的上游,并显微注射到卵母细胞和受精卵中。通过CAT分析和引物延伸来确定活性。c-myc启动子能非常有效地驱动转录,一个截短的启动子-158/+46基本上保留了全部活性。该区域包含一个重叠的E2F/SP1位点和两个与小鼠c-myc基因中发现的串联Sp1位点同源的位点。内部缺失表明,这两个元件在驱动CAT表达方面在卵母细胞中具有同等活性。生发泡提取物含有一种对Sp1共有序列具有特异性的DNA结合活性,但对E2F位点没有活性。数据表明,非洲爪蟾卵母细胞中内源性c-myc基因的高转录水平是由Sp1或一种相关转录因子介导的。在胚胎中出现了不同的机制,Sp1结合位点的功能作用似乎不那么重要。

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