Formation of the transcription initiation complex on mammalian rDNA.
- 1 October 1986
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 6 (10) , 3418-3427
- https://doi.org/10.1128/mcb.6.10.3418
Abstract
Steps for the formation of transcription initiation complex on the human rRNA gene (rDNA) in vitro were analyzed with partially purified transcription factors and RNA polymerase I. The reaction requires at least two factors besides RNA polymerase I for maximal efficiency. Preincubation and short-pulse analyses of the accurate transcripts revealed the following steps. First, the species-dependent factor, designated TFID, bound to the rDNA template, forming a preinitiation complex (PIC-1) which was resistant to a moderate concentration (0.015 to 0.02%) of Sarkosyl. Other factors, designated TFIA and RNA polymerase I, were then added to convert it to the final preinitiation complex PIC-3. This complex incorporated the first two nucleoside triphosphates of the starting site to complete the initiation complex (IC), which was resistant to a high concentration (0.2%) of Sarkosyl. Binding of TFID was rate limiting in the overall initiation reaction in vitro. Together with the kinetics of incorporation, the results are interpreted to mean that TFID, once bound, remains complexed with rDNA together with TFIA as the PIC-2 for many rounds of transcription by RNA polymerase I. Thus, the formation of PIC-2 may be a prerequisite for the stable opening of rDNA for transcription in vivo.This publication has 40 references indexed in Scilit:
- Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box regionCell, 1985
- Formation of a rate-limiting intermediate in 5S RNA gene transcriptionCell, 1985
- A drosophila RNA polymerase II transcription factor binds to the regulatory site of an hsp 70 geneCell, 1984
- Transcription of Class III Genes: Formation of Preinitiation ComplexesScience, 1983
- Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase IICell, 1983
- Formation of stable preinitiation complexes between eukaryotic class B transcription factors and promoter sequencesNature, 1983
- Specific interaction of a purified transcription factor with an internal control region of 5S RNA genesCell, 1980
- Selective and accurate initiation of transcription at the ad2 major late promotor in a soluble system dependent on purified rna polymerase ii and dnaCell, 1979
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- Isolation of DNA segments containing promoters from bacteriophage T3 DNABiochemistry, 1974