title: Loss of function and inhibitory effects of human CSX/NKX2.5 homeoprotein mutations associated with congenital heart disease.
abstract: CSX/NKX2.5 is an evolutionarily conserved homeodomain-containing (HD-containing) transcription factor that is essential for early cardiac development. Recently, ten different heterozygous CSX/NKX2.5 mutations were found in patients with congenital heart defects that are transmitted in an autosomal dominant fashion. To determine the consequence of these mutations, we analyzed nuclear localization, DNA binding, transcriptional activation, and dimerization of mutant CSX/NKX2.5 proteins. All mutant proteins were translated and located to the nucleus, except one splice-donor site mutant whose protein did not accumulate in the cell. All mutants that had truncation or missense mutations in the HD had severely reduced DNA binding activity and little or no transcriptional activation function. In contrast, mutants with intact HDs exhibit normal DNA binding to the monomeric binding site but had three- to ninefold reduction in DNA binding to the dimeric binding sites. HD missense mutations that preserved homodimerization ability inhibited the activation of atrial natriuretic factor by wild-type CSX/NKX2.5. Although our studies do not characterize the genotype-phenotype relationship of the ten human mutations, they identify specific abnormalities of CSX/NKX2.5 function essential for transactivation of target genes.
authors.affiliation: Cardiovascular Division, Beth Israel Deaconess Medical Center, Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: H
authors.initials: H
authors.lastname: Kasahara
authors.forename: B
authors.initials: B
authors.lastname: Lee
authors.forename: J J
authors.initials: JJ
authors.lastname: Schott
authors.forename: D W
authors.initials: DW
authors.lastname: Benson
authors.forename: J G
authors.initials: JG
authors.lastname: Seidman
authors.forename: C E
authors.initials: CE
authors.lastname: Seidman
authors.forename: S
authors.initials: S
authors.lastname: Izumo
mesh_terms: D002451:Cell Compartmentation; D019281:Dimerization; D006327:Heart Block / Q000235:genetics*; D006331:Heart Diseases / Q000151:congenital*; D006344:Heart Septal Defects, Atrial / Q000235:genetics*; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D009154:Mutation; D020125:Mutation, Missense; D009687:Nuclear Proteins / Q000235:genetics; D011485:Protein Binding; D014176:Protein Biosynthesis; D012326:RNA Splicing; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation; D029867:Xenopus Proteins
chemical_list: D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C095552:NKX2-5 protein, human; D009687:Nuclear Proteins; D014157:Transcription Factors; D029867:Xenopus Proteins