Linked Life Data

20450920

Source:http://linkedlifedata.com/resource/pubmed/id/20450920

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2010-6-14
pubmed:databankReference
pubmed:abstractText
TBX5, a member of the T-box transcription factor family, plays an important role in heart and limb development. More than 60 single point or deletion mutations of human TBX5 are associated with Holt-Oram syndrome that manifests itself as heart and limb malformations in 1 out of 100,000 live births. The majority of these mutations are located in the TBX5 T-box domain. We solved the crystal structures of the human TBX5 T-box domain in its DNA-unbound form and in complex with a natural DNA target site allowing for the first time the comparison between unbound and DNA-bound forms. Our analysis identifies a 3(10)-helix at the C-terminus of the T-box domain as an inducible recognition element, critically required for the interaction with DNA, as it only forms upon DNA binding and is unstructured in the DNA-unbound form. Using circular dichroism, we characterized the thermal stability of six TBX5 mutants containing single point mutations in the T-box domain (M74V, G80R, W121G, G169R, T223M, and R237W) and compared them with wild-type protein. Mutants G80R and W121G show drastically reduced thermal stability, while the other mutants only show a marginal stability decrease. For all TBX5 mutants, binding affinities to specific and nonspecific DNA sequences were determined using isothermal titration calorimetry. All TBX5 mutants show reduced binding affinities to a specific DNA target site, although to various degrees. Interestingly, all tested TBX5 mutants differ in their ability to bind unspecific DNA, indicating that both sequence-specific and unspecific binding might contribute to the misregulation of target gene expression.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
1089-8638
pubmed:author
pubmed:copyrightInfo
Copyright 2010 Elsevier Ltd. All rights reserved.
pubmed:issnType
Electronic
pubmed:day
2
pubmed:volume
400
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
71-81
pubmed:meshHeading
pubmed-meshheading:20450920-Abnormalities, Multiple, pubmed-meshheading:20450920-Amino Acid Sequence, pubmed-meshheading:20450920-Animals, pubmed-meshheading:20450920-Base Sequence, pubmed-meshheading:20450920-Circular Dichroism, pubmed-meshheading:20450920-DNA, pubmed-meshheading:20450920-Fetal Proteins, pubmed-meshheading:20450920-Heart Defects, Congenital, pubmed-meshheading:20450920-Humans, pubmed-meshheading:20450920-Limb Deformities, Congenital, pubmed-meshheading:20450920-Models, Molecular, pubmed-meshheading:20450920-Molecular Sequence Data, pubmed-meshheading:20450920-Nucleic Acid Conformation, pubmed-meshheading:20450920-Point Mutation, pubmed-meshheading:20450920-Protein Structure, Tertiary, pubmed-meshheading:20450920-Sequence Alignment, pubmed-meshheading:20450920-T-Box Domain Proteins
pubmed:year
2010
pubmed:articleTitle
Structural basis of TBX5-DNA recognition: the T-box domain in its DNA-bound and -unbound form.
pubmed:affiliation
European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't