Effect of the non-conserved N-terminus on the DNA binding activity of the yeast TATA binding protein

We have studied the DNA binding activity of recombinant yeast TATA Binding Protein (TBP) with particular interest in the role played by the nonconserved N-terminal domain. By comparing the DNA binding activity of wild type yeast TBP with a mutant form of TBP that lacks the non-conserved N-terminal d...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 1993-04, Vol.21 (8), p.1789-1796
Main Authors: Kuddus, R, Schmidt, M.C
Format: Article
Language:English
Subjects:
amino acid sequences
Base Sequence
binding sites
Biological and medical sciences
DNA, Fungal - chemistry
DNA, Fungal - metabolism
DNA-binding proteins
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Fundamental and applied biological sciences. Psychology
Molecular and cellular biology
molecular conformation
Molecular genetics
Molecular Sequence Data
Mutation
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Solutions
TATA Box
tata box element
TATA-Box Binding Protein
Transcription Factors - chemistry
Transcription Factors - genetics
Transcription Factors - metabolism
Transcription. Transcription factor. Splicing. Rna processing
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We have studied the DNA binding activity of recombinant yeast TATA Binding Protein (TBP) with particular interest in the role played by the nonconserved N-terminal domain. By comparing the DNA binding activity of wild type yeast TBP with a mutant form of TBP that lacks the non-conserved N-terminal domain (TBP delta 57), we have determined that the N-terminus of TBP alters both the shape and the stability of the TBP-DNA complex. Measurements of the DNA bending angle indicate that the N-terminus enhances the bending of the DNA that is induced by TBP binding and greatly destabilizes the TBP-DNA complex during native gel electrophoresis. In solution, the N-terminus has only a slight effect on the equilibrium dissociation constant and the dissociation rate constant. However, the N-terminal domain reduces the association rate constant in a temperature dependent manner and increases the apparent activation energy of the TBP-DNA complex formation by 3 kcal/mole. These data suggest that a conformational change involving the N-terminus of TBP may be one of the isomerization steps in the formation of a stable TBP-DNA complex.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/21.8.1789