Monitoring the interaction between DNA and a transcription factor (MEF2A) using fluorescence correlation spectroscopy

详细信息    查看全文

• 作者：Guillaume Octobre ; Claudie Lemercier ; Saadi Khochbin ; Michel Robert-Nicoud and Catherine Souchier
• 关键词：DNA-binding ; MEF2 ; MADS box ; Fluorescence correlation spectroscopy
  Mots-clé ; s ; Interaction ADN-proté ; ine ; MEF2 ; MADS box ; Spectroscopie à ; corré ; lation de fluorescence
• 刊名：Comptes Rendus Biologies
• 出版年：2005
• 出版时间：December 2005
• 年：2005
• 卷：328
• 期：12
• 页码：1033-1040
• 全文大小：319 K

文摘

Fluorescence correlation spectroscopy (FCS) is an analytical method that allows distinguishing different populations of fluorescent probes in solution and provides data on their concentrations and their diffusion coefficients. FCS was used to characterize the interaction of the transcription factor (MEF2A) with its DNA target sequence. The myocyte enhancer factor 2 (MEF2) belongs to the MADS-box family and activates transcription of numerous muscle genes during myogenesis. Measurements were made using TAMRA-labelled oligonucleotide duplexes derived from a wild type (WT) or a mutated MEF2 target gene. Binding of the protein to the WT DNA resulted in significant changes of the diffusion. Specificity of the interaction was confirmed using the mutated DNA. Bound to free probe ratios were determined at different MEF2A concentrations and the apparent equilibrium dissociation constant K_D for the full-length MEF2A was estimated. To cite this article: G. Octobre et al., C. R. Biologies 328 (2005).