Factors involved in specific transcription by mammalian RNA polymerase II: purification and characterization of general transcription factor TFIIE.

RAP30/74 is a human general transcription factor that binds to RNA polymerase II and is required for initiation of transcription in vitro regardless of whether the promoter has a recognizable TATA box (Z. F. Burton, M. Killeen, M. Sopta, L. G. Ortolan, and J. F. Greenblatt, Mol. Cell. Biol. 8:1602-1613, 1988). Part of the amino acid sequence of RAP30, the small subunit of RAP30/74, has limited homology with part of Escherichia coli sigma 70 (M. Sopta, Z. F. Burton, and J. Greenblatt, Nature (London) 341:410-414, 1989). To determine which sigmalike activities of RAP30/74 could be attributed to RAP30, we purified human RAP30 and a RAP30-glutathione-S-transferase fusion protein that had been produced in E. coli. Bacterially produced RAP30 bound to RNA polymerase II in the absence of RAP74. Both partially purified natural RAP30/74 and recombinant RAP30 prevented RNA polymerase II from binding nonspecifically to DNA. In addition, nonspecific transcription by RNA polymerase II was greatly inhibited by RAP30-glutathione-S-transferase. DNA-bound RNA polymerase II could be removed from DNA by partially purified RAP30/74 but not by bacterially expressed RAP30. Thus, the ability of RAP30/74 to recruit RNA polymerase II to a promoter-bound preinitiation complex may be an indirect consequence of its ability to suppress nonspecific binding of RNA polymerase II to DNA.

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DNA-dependent RNA Polymerase from Ehrllch Ascites Tumor CellsI. Partial Purification and Characterization of RNA Polymerase II

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a130091 ◽

1973 ◽

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Tumor Cells ◽

Partial Purification ◽

Ascites Tumor ◽

Purification And Characterization

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Interaction of the TFIIB zinc ribbon with RNA polymerase II

Biochemical Society Transactions ◽

10.1042/bst0360595 ◽

2008 ◽

Vol 36 (4) ◽

pp. 595-598 ◽

Cited By ~ 6

Author(s):

Laura M. Elsby ◽

Stefan G.E. Roberts

Keyword(s):

Transcription Factor ◽

Transcription Factors ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Structural Models ◽

Initiation Complex ◽

General Transcription Factors ◽

Transcription Factor Iib ◽

General Transcription Factor ◽

Zinc Ribbon

Transcription by RNA polymerase II requires the assembly of the general transcription factors at the promoter to form a pre-initiation complex. The general transcription factor TF (transcription factor) IIB plays a central role in the assembly of the pre-initiation complex, providing a bridge between promoter-bound TFIID and RNA polymerase II/TFIIF. We have characterized a series of TFIIB mutants in their ability to support transcription and recruit RNA polymerase II to the promoter. Our analyses identify several residues within the TFIIB zinc ribbon that are required for RNA polymerase II assembly. Using the structural models of TFIIB, we describe the interface between the TFIIB zinc ribbon region and RNA polymerase II.

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A Fully Recombinant System for Activator-dependent Archaeal Transcription

Journal of Biological Chemistry ◽

10.1074/jbc.c400446200 ◽

2004 ◽

Vol 279 (50) ◽

pp. 51719-51721 ◽

Cited By ~ 51

Author(s):

Mohamed Ouhammouch ◽

Finn Werner ◽

Robert O. J. Weinzierl ◽

E. Peter Geiduschek

Keyword(s):

Transcription Factor ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Regulators ◽

The Core ◽

General Transcription Factor ◽

Core Components ◽

Archaeal Transcription ◽

Bacterial Type

The core components of the archaeal transcription apparatus closely resemble those of eukaryotic RNA polymerase II, while the DNA-binding transcriptional regulators are predominantly of bacterial type. Here we report the construction of an entirely recombinant system for positively regulated archaeal transcription. By omitting individual subunits, or sets of subunits, from thein vitroassembly of the 12-subunit RNA polymerase from the hyperthermophileMethanocaldococcus jannaschii, we describe a functional dissection of this RNA polymerase II-like enzyme, and its interactions with the general transcription factor TFE, as well as with the transcriptional activator Ptr2.

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