Visualization of bHLH transcription factor interactions in living mammalian cell nuclei and developing chicken neural tube by FRET

	Visualization of bHLH transcription factor interactions in living mammalian cell nuclei and developing chicken neural tube by FRET
	Wang, C; Bian, W; Xia, CH; Zhang, T; Guillemot, F; Jing, NH
刊名	CELL RESEARCH
	2006
卷号	16 期号:6 页码:585-598
关键词	bHLH confocal microscopy chick embryo neural tube FRET in ovo electroporation photobleach
通讯作者	Jing, NH (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Stem Cell Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,njing@sibs.ac.cn
英文摘要	Members of the basic helix-loop-helix (bHLH) gene family play important roles in vertebrate neurogenesis. In this study, confocal microscopy-based fluorescence resonance energy transfer (FRET) is used to monitor bHLH protein-protein interactions under various physiological conditions. Tissue-specific bHLH activators, NeuroD I, Mash I, Neurogenin I (Ngn1), Neurogenin2 (Ngn2), and ubiquitous expressed E47 protein are tagged with enhanced yellow fluorescence protein (EYFP) and enhanced cyan fluorescence protein (ECFP), respectively. The subcellular localization and mobility of bHLH fusion proteins are examined in HEK293 cells. By transient transfection and in ovo electroporation, four pairs of tissue-specific bHLH activators and E47 protein are over-expressed in HEK293 cells and developing chick embryo neural tube. With the acceptor photobleaching method, FRET could be detected between these bHLH protein pairs in the nuclei of transfected cells and developing neural tubes. Mash1/E47 and Ngn2/E47 FRET pairs show higher FRET efficiencies in the medial and the lateral half of chick embryo neural tube, respectively. It suggests that these bHLH protein pairs formed functional DNA-protein complexes with regulatory elements of their downstream target genes in the specific regions. This work will help one understand the behaviours of bHLH factors in vivo.
学科主题	Cell Biology
类目[WOS]	Cell Biology
关键词[WOS]	RESONANCE ENERGY-TRANSFER ; LOOP-HELIX PROTEIN ; DNA-BINDING ; TRANSFER MICROSCOPY ; PRONEURAL GENES ; SENSORY NEURONS ; NERVOUS-SYSTEM ; STEM-CELLS ; NEUROGENESIS ; EXPRESSION
收录类别	SCI
语种	英语
WOS记录号	WOS:000239296000008
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1730]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Wang, C,Bian, W,Xia, CH,et al. Visualization of bHLH transcription factor interactions in living mammalian cell nuclei and developing chicken neural tube by FRET[J]. CELL RESEARCH,2006,16(6):585-598.
APA	Wang, C,Bian, W,Xia, CH,Zhang, T,Guillemot, F,&Jing, NH.(2006).Visualization of bHLH transcription factor interactions in living mammalian cell nuclei and developing chicken neural tube by FRET.CELL RESEARCH,16(6),585-598.
MLA	Wang, C,et al."Visualization of bHLH transcription factor interactions in living mammalian cell nuclei and developing chicken neural tube by FRET".CELL RESEARCH 16.6(2006):585-598.