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Related Concept Videos

Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Transcription Elongation Factors02:35

Transcription Elongation Factors

Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
Transcription Elongation Factors02:35

Transcription Elongation Factors

Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
General Transcription Factors01:30

General Transcription Factors

Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
Bacterial Transcription01:53

Bacterial Transcription

RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:

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Updated: May 27, 2026

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events
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Published on: May 13, 2019

Efficient transcription by RNA polymerase I using recombinant core factor.

Gregory J Bedwell1, Francis D Appling, Susan J Anderson

  • 1Department of Biochemistry and Molecular Genetics, University of Alabama-Birmingham, 720 20th Street South, Birmingham, AL 35294, USA.

Gene
|November 19, 2011
PubMed
Summary

Researchers developed a new method to express and purify a key complex for ribosomal DNA transcription. This recombinant core factor is more active and aids in understanding RNA polymerase I transcription regulation.

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Last Updated: May 27, 2026

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Ribosome biogenesis is critical for cell proliferation and relies on ribosomal DNA (rDNA) transcription by RNA polymerase I.
  • Understanding the molecular mechanisms controlling RNA polymerase I transcription is essential for defining cell proliferation regulation.
  • Existing biochemical assays for RNA polymerase I transcription factors are limited by the low abundance and complex purification of essential components.

Purpose of the Study:

  • To develop an efficient method for expressing and purifying the three-subunit core factor complex for RNA polymerase I transcription.
  • To compare the activity of the recombinant core factor with yeast-derived core factor in transcription assays.
  • To utilize the recombinant core factor to investigate the role of TATA-binding protein in RNA polymerase I transcription.

Main Methods:

  • Expression and purification of the three-subunit core factor complex from Escherichia coli.
  • In vitro transcription assays using purified recombinant core factor.
  • Biochemical analyses to compare recombinant and yeast-derived core factor activity.
  • Utilizing recombinant core factor to test models of TATA-binding protein function.

Main Results:

  • A novel method for expressing and purifying the three-subunit core factor complex from E. coli was established.
  • The recombinant core factor exhibited higher activity in RNA polymerase I transcription assays compared to yeast-derived core factor.
  • The recombinant core factor enabled differentiation between two models regarding the TATA-binding protein's role in RNA polymerase I transcription.

Conclusions:

  • The developed recombinant expression and purification method provides a more active and accessible core factor for studying RNA polymerase I transcription.
  • This advancement facilitates a deeper understanding of the regulation of eukaryotic ribosome biogenesis and cell proliferation.
  • The recombinant core factor is a valuable tool for dissecting the precise roles of transcription factors, including TATA-binding protein, in rDNA transcription.