SEQUENCE AND FACTOR REQUIREMENTS FOR FAITHFUL INVITRO TRANSCRIPTION OF HUMAN 7SL DNA

被引：28

作者：

BREDOW, S ^{[1
]}

KLEINERT, H ^{[1
]}

BENECKE, BJ ^{[1
]}

机构：

[1] RUHR UNIV BOCHUM,DEPT BIOCHEM NC6,W-4630 BOCHUM 1,GERMANY

来源：

GENE | 1990年 / 86卷 / 02期

关键词：

deletion mutants; footprinting; recombinant DNA; RNA polymerase III promoter; template-exclusion assay; transcription factors;

D O I：

10.1016/0378-1119(90)90282-V

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have analysed the transcription of a functional 7SL gene by RNA polymerase III (RNAPIII) in S100 extracts in vitro. Accurate and efficient synthesis of 7S L RNA depends on the presence of (i) an upstream sequence and (ii) an internal promoter element located within the first 22 bp of the gene. These findings were substantiated by DNase I footprinting. Mutations of the internal promoter identified the doublet CG [nucleotide (nt) +15/+16] outside the A-box homologue (nt +15/ to +14) as being essential for both proper promoter function in the vitro transcription assay and competition in the template-exclusion assay. Fractionation of S100 extracts identified two fractions required in addition to RNAPIII for faithful transcription of the gene. Each of these two fractions gave rise to one of two footprints observed in DNase I protection experiments, indicating that at least two DNA-binding factors are involved. © 1990.

引用

页码：217 / 225

页数：9

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