EGFP-tagged core and linker histones diffuse via distinct mechanisms within living cells.
The effect of chromatin organization on EGFP-tagged histone protein dynamics within the cell nucleus has been probed using fluorescence correlation and recovery measurements on single living HeLa cells. Our studies reveal that free fraction of core-particle histones exist as multimers within the cell nucleus whereas the linker histones exist in monomeric forms. The multimeric state of core histones is found to be invariant across mammalian and polytene chromosomes and this is ATP dependent. In contrast, the dynamics of the linker histones exhibits two distinct diffusion timescales corresponding to its transient binding and unbinding to chromatin governed by the tail domain residues. Under conditions of chromatin condensation induced by apoptosis, the free multimeric fraction of core histones is found to become immobile, while the monomeric linker histone mobility is partially reduced. In addition, we observe differences in nuclear colocalization of linker and core particle histones. These results are validated through Brownian dynamics simulation of core and linker histone mobility. Our findings provide a framework to understand the coupling between the state of chromatin assembly and histone protein dynamics that is central to accessing regulatory sites on the genome.
Bhattacharya
Dipanjan
D
National Center for Biological Sciences, TIFR, Bangalore-560065, India.
Mazumder
Aprotim
A
Miriam
S Annie
SA
Shivashankar
G V
GV
D000255:Adenosine Triphosphate / Q000172:deficiency; D017209:Apoptosis; D002467:Cell Nucleus / Q000502:physiology*; D002843:Chromatin / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D004058:Diffusion; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006367:HeLa Cells; D006657:Histones / Q000502:physiology*; D006801:Humans; D021381:Protein Transport; D013050:Spectrometry, Fluorescence
D002843:Chromatin; D006657:Histones; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate