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小鼠DNA聚合酶β基因启动子:精细定位及类Sp1小鼠转录因子在其功能中的作用

Mouse DNA polymerase beta gene promoter: fine mapping and involvement of Sp1-like mouse transcription factor in its function.

作者信息

Yamaguchi M, Hayashi Y, Matsukage A

机构信息

Laboratory of Cell Biology, Aichi Cancer Center Research Institute, Nagoya, Japan.

出版信息

Nucleic Acids Res. 1988 Sep 26;16(18):8773-87. doi: 10.1093/nar/16.18.8773.

Abstract

The promoter of mouse DNA polymerase beta gene was analyzed by combining 5'-upstream region of this gene with chloramphenicol acetyltransferase (CAT) gene and by introduction of the recombinant plasmid DNA into mouse NIH/3T3 cells. Serial deletion of the mouse DNA sequence revealed that the promoter function resides within a 33 base pair region from the nucleotide position -48 to -15 with respect to the transcription initiation site, and is highly active without enhancer sequence. The promoter region was separated into two subregions: one (-48 to -35) contains a GC-box and the other contains a 10 base pair palindrome, whose sequence is similar to one of promoter consensus sequences found in a number of promoters including adenovirus promoters. The DNA polymerase beta promoter-directed CAT expression was competitively inhibited by the simultaneous transfection of plasmid DNA containing SV40 early promoter sequence. The viral sequences which are competitive to the GC-box of DNA polymerase beta gene promoter were the GC-boxes of SV40 promoter. Therefore, it is concluded that transcription of mouse DNA polymerase beta gene is regulated by mouse trans-acting factors equivalent to human Sp1 which is known to be trans-acting protein factor acting on SV40 GC-box sequences.

摘要

通过将小鼠DNA聚合酶β基因的5'上游区域与氯霉素乙酰转移酶(CAT)基因相结合,并将重组质粒DNA导入小鼠NIH/3T3细胞,对小鼠DNA聚合酶β基因的启动子进行了分析。对小鼠DNA序列的系列缺失分析表明,启动子功能存在于相对于转录起始位点从核苷酸位置-48至-15的33个碱基对区域内,并且在没有增强子序列的情况下具有高活性。启动子区域被分为两个子区域:一个(-48至-35)包含一个GC盒,另一个包含一个1碱基对的回文序列,其序列类似于在包括腺病毒启动子在内的许多启动子中发现的启动子共有序列之一。同时转染含有SV40早期启动子序列的质粒DNA可竞争性抑制DNA聚合酶β启动子指导的CAT表达。与DNA聚合酶β基因启动子的GC盒竞争的病毒序列是SV40启动子的GC盒。因此,可以得出结论,小鼠DNA聚合酶β基因的转录受与人类Sp1等效的小鼠反式作用因子调节,已知人类Sp1是作用于SV40 GC盒序列的反式作用蛋白因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1436/338634/ed9b806f4769/nar00160-0037-a.jpg

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