Trichostatin A-induced histone acetylation causes decondensation of interphase chromatin.
The effect of trichostatin A (TSA)-induced histone acetylation on the interphase chromatin structure was visualized in vivo with a HeLa cell line stably expressing histone H2A, which was fused to enhanced yellow fluorescent protein. The globally increased histone acetylation caused a reversible decondensation of dense chromatin regions and led to a more homogeneous distribution. These structural changes were quantified by image correlation spectroscopy and by spatially resolved scaling analysis. The image analysis revealed that a chromatin reorganization on a length scale from 200 nm to >1 microm was induced consistent with the opening of condensed chromatin domains containing several Mb of DNA. The observed conformation changes could be assigned to the folding of chromatin during G1 phase by characterizing the effect of TSA on cell cycle progression and developing a protocol that allowed the identification of G1 phase cells on microscope coverslips. An analysis by flow cytometry showed that the addition of TSA led to a significant arrest of cells in S phase and induced apoptosis. The concentration dependence of both processes was studied.
Kirchhoff-Institut für Physik, AG Molekulare Biophysik, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 227, 69120 Heidelberg, Germany.
Katalin Fejes
KF
Tóth
Tobias A
TA
Knoch
Malte
M
Wachsmuth
Monika
M
Frank-Stöhr
Michael
M
Stöhr
Christian P
CP
Bacher
Gabriele
G
Müller
Karsten
K
Rippe
D000107:Acetylation / Q000187:drug effects; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D002843:Chromatin / Q000187:drug effects* / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000187:drug effects / Q000378:metabolism; D017709:Fractals; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology; D018816:Genes, cdc / Q000187:drug effects / Q000502:physiology; D006367:HeLa Cells; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007091:Image Processing, Computer-Assisted; D007399:Interphase / Q000187:drug effects / Q000502:physiology; D046032:Microscopy, Energy-Filtering Transmission Electron; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects / Q000502:physiology
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D006877:Hydroxamic Acids; D011500:Protein Synthesis Inhibitors; C051886:chromosome decondensation factors; C012589:trichostatin A; D006655:Histone Deacetylases