Mutations within helix I of twist1 result in distinct limb defects and variation of DNA binding affinities

Beth A. Firulli, Bradley A. Redick, Simon Conway, Anthony Firulli

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Twist1 is a basic helix-loop-helix (bHLH) factor that plays an important role in limb development. Haploinsufficiency of Twist1 results in polydactyly via the inability of Twist1 to antagonistically regulate the related factor Hand2. The mechanism modulating Twist1-Hand2 antagonism is via phosphoregulation of conserved threonine and serine residues in helix I of the bHLH domain. Phosphoregulation alters the dimerization affinities for both proteins. Here we show that the expression of Twist1 and Twist1 phosphoregulation mutants results in distinct limb phenotypes in mice. In addition to dimer regulation, Twist1 phosphoregulation affects the DNA binding affinities of Twist1 in a partner-dependent and cis-element-dependent manner. In order to gain a better understanding of the specific Twist1 transcriptional complexes that function during limb morphogensis, we employ a series of Twist1-tethered dimers that include the known Twist1 partners, E12 and Hand2, as well as a tethered Twist1 homodimer. We show that these dimers behave in a manner similar to monomerically expressed bHLH factors and result in distinct limb phenotypes that correlate well with those observed from the limb expression of Twist1 and Twist1 phosphoregulation mutants. Taken together, this study shows that the Twist1 dimer affinity for a given partner can modulate the DNA binding affinity and that Twist1 dimer choice determines phenotypic outcome during limb development.

Original languageEnglish
Pages (from-to)27536-27546
Number of pages11
JournalJournal of Biological Chemistry
Volume282
Issue number37
DOIs
StatePublished - Sep 14 2007

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Dimers
Extremities
Defects
Mutation
DNA
Helix-Loop-Helix Motifs
Dimerization
Polydactyly
Phenotype
Haploinsufficiency
Threonine
Serine
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mutations within helix I of twist1 result in distinct limb defects and variation of DNA binding affinities. / Firulli, Beth A.; Redick, Bradley A.; Conway, Simon; Firulli, Anthony.

In: Journal of Biological Chemistry, Vol. 282, No. 37, 14.09.2007, p. 27536-27546.

Research output: Contribution to journalArticle

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