title: Multiscale analysis of dynamics and interactions of heterochromatin protein 1 by fluorescence fluctuation microscopy.
abstract: Heterochromatin protein 1 (HP1) is a central factor in establishing and maintaining the repressive heterochromatin state. To elucidate its mobility and interactions, we conducted a comprehensive analysis on different time and length scales by fluorescence fluctuation microscopy in mouse cell lines. The local mobility of HP1alpha and HP1beta was investigated in densely packed pericentric heterochromatin foci and compared with other bona fide euchromatin regions of the nucleus by fluorescence bleaching and correlation methods. A quantitative description of HP1alpha/beta in terms of its concentration, diffusion coefficient, kinetic binding, and dissociation rate constants was derived. Three distinct classes of chromatin-binding sites with average residence times t(res) <or= 0.2 s (class I, dominant in euchromatin), 7 s (class II, dominant in heterochromatin), and approximately 2 min (class III, only in heterochromatin) were identified. HP1 was present at low micromolar concentrations at heterochromatin foci, and required histone H3 lysine 9 methylases Suv39h1/2 for two- to fourfold enrichment at these sites. These findings impose a number of constraints for the mechanism by which HP1 is able to maintain a heterochromatin state.
authors: Deutsches Krebsforschungszentrum and BioQuant, Research Group Genome Organization and Function, Heidelberg, Germany.
authors: Katharina P
authors: KP
authors: Müller
authors: Fabian
authors: F
authors: Erdel
authors: Maïwen
authors: M
authors: Caudron-Herger
authors: Caroline
authors: C
authors: Marth
authors: Barna D
authors: BD
authors: Fodor
authors: Mario
authors: M
authors: Richter
authors: Manuela
authors: M
authors: Scaranaro
authors: Joël
authors: J
authors: Beaudouin
authors: Malte
authors: M
authors: Wachsmuth
authors: Karsten
authors: K
authors: Rippe
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004058:Diffusion; D044127:Epigenesis, Genetic; D036681:Fluorescence Recovery After Photobleaching; D006570:Heterochromatin / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008856:Microscopy, Fluorescence; D009068:Movement; D021381:Protein Transport; D013050:Spectrometry, Fluorescence
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D006570:Heterochromatin; D000090266:Chromobox Protein Homolog 5; D011495:Histone-Lysine N-Methyltransferase