TBX5-NKX2

Crystallographic and biochemical studies of the human TBX5-Nkx2-5-DNA complex

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 Obiettivo del progetto (Objective)

'Holt Oram syndrome (HOS) is a heart and limb syndrome, which manifests itself in 1 of 100000 live births. HOS syndrome is an autosomal-dominant disease, where forelimb and cardiac congenital abnormalities are observed. The syndrome is connected to mutations in the T-box transcription factor TBX5 gene. Similar symptoms are observed when mutations in the homeobox transcription factor human Nkx2-5 are present. TBX5 and Nkx2-5 interact with each other and synergistically activate the promoters of the cardiac-specific natriuretic peptide precursor type A (ANF) and connexin (cx40). The project goal is the biochemical characterization and crystal structure solution of the ternary TBX5-Nkx2-5-DNA complex. The crystal structure of the ternary complex will allow a comprehensive structural analysis of the interaction mechanism between TBX5 and Nkx2-5 and their DNA binding sites and will elucidate the synergistic promoter activation of TBX5 and Nkx2-5 on a molecular level. Such insight is of broad interest for cardiovascular biologist because it opens the ultimate perspective to use this knowledge for influencing cardiac growth and for cardiac regeneration. The results of this project will contribute to the European quality and competitiveness in cardio vascular research. The project uses the full portfolio of structural biological research and requires interdisciplinary knowledge in molecular biology, biochemistry, biophysics, protein crystallography and bioinformatical techniques. Thus, the researcher has the possibility to gain skills in all techniques necessary for doing successful research in structural biology. In addition, the multidisciplinary environment at the EMBL will give the researcher the possibility to interact and communicate with scientists in fields other than crystallography. All mentioned points will allow the scientist to gain professional maturity and to develop an independent research career.'

Introduzione (Teaser)

An EU-funded research team successfully characterised the complex formed by two transcription factors implicated in Holt Oram Syndrome. Advances in this area have the potential to help future heart patients.

Descrizione progetto (Article)

Holt Oram Syndrome (HOS) is a heart and limb syndrome, which manifests itself in 1 in 100,000 live births. The syndrome is connected to mutations in the transcription factor TBX5 gene. Similar symptoms to HOS are observed when mutations in the human transcription factor Nkx2-5 are present. TBX5 and Nkx2-5 play central roles in cardiac development and on interaction synergistically activate the promoters of the cardiac-specific peptide precursor type A (ANF) and connexin (cx40).

The goal of the 'Crystallographic and biochemical studies of the human TBX5-Nkx2-5-DNA complex' (TBX5-NKX2) project was to characterise the TBX5-Nkx2-5-DNA complex and determine its crystal structure. The project applied structural biology techniques including cloning, protein expression and purification, protein/DNA interaction studies, crystallisation and crystallography to characterise the complex.

Project partners made solid progress and insights into the TBX5 specific promoter recognition were obtained together with significant structural and biophysical data on the complex. The full crystal structure will be published in a leading academic journal in the near future. Knowledge of the crystal structure allows for a comprehensive structural analysis of the interaction mechanisms between TBX5 and Nkx2-5 and their DNA binding sites. It also promises to give a better understanding of the synergistic promoter activation of TBX5 and Nkx2-5 at a molecular level.

Gaining these insights is of broad interest for cardiovascular biology because this knowledge can be used for influencing cardiac growth and in cardiac regeneration in patients.