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Related Concept Videos

Transcription Factors02:16

Transcription Factors

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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...
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Protein Complexes with Interchangeable Parts01:57

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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
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Transfer RNA Synthesis02:36

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One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
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Eukaryotic Transcription Inhibitors01:52

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Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
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Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

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Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
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General Transcription Factors01:30

General Transcription Factors

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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...
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Related Experiment Video

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A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies
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A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

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Yeast and human TFIID with altered DNA-binding specificity for TATA elements.

M Strubin1, K Struhl

  • 1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.

Cell
|February 21, 1992
PubMed
Summary

Researchers engineered a yeast TFIID protein to recognize mutated TATA elements, crucial for RNA polymerase II transcription. This modification also enabled human TFIID to function in yeast, revealing key interactions in gene regulation.

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Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences
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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Protein Engineering

Background:

  • TFIID is a conserved transcription factor essential for RNA polymerase II activity.
  • TFIID specifically binds to the TATA element (consensus TATAAA) in gene promoters.
  • Understanding TFIID-TATA interactions is key to deciphering transcription regulation.

Purpose of the Study:

  • To isolate and characterize a yeast TFIID derivative with altered TATA element specificity.
  • To identify the specific amino acid changes responsible for the altered TFIID binding.
  • To assess the functional conservation of these changes in human TFIID within a yeast system.

Main Methods:

  • Genetic selection was employed to isolate yeast TFIID mutants with altered promoter recognition.
  • Biochemical analysis was used to confirm the binding specificity of the mutant TFIID.
  • Site-directed mutagenesis was performed to introduce specific amino acid substitutions into yeast and human TFIID.

Main Results:

  • An altered specificity yeast TFIID derivative was isolated, capable of binding mutated TATA elements (TGTAAA).
  • Three amino acid substitutions in TFIID, two being critical, conferred the altered binding specificity.
  • Human TFIID with analogous substitutions supported transcription from a TGTAAA promoter in yeast.

Conclusions:

  • A specific surface of TFIID directly interacts with the TATA element.
  • Amino acid substitutions can alter TFIID specificity for TATA box sequences.
  • Human TFIID retains functional compatibility with yeast transcription machinery and can be engineered for altered specificity.