Chromatin structural transitions following histone H1 displacement by phosphatidylserine vesicles and low pH treatment. A multiparametric analysis involving flow cytometry, electron microscopy, and nuclease digestion.
We describe several morphological and functional modifications in isolated rat liver nuclei incubated in the presence of phosphatidylserine (PS) multilamellar vesicles (MLV). These effects, which occur through the release of histone H1, induce chromatin decondensation, as shown by electron microscopy and nuclease digestion. Flow cytometry was employed to monitor these changes in chromatin structure in isolated nuclei by means of perpendicular light scatter (PLS) and fluorescence signals. Chromatin decondensation induced by PS or by low pH treatment was accompanied by an increase in perpendicular light scatter and by less efficient binding of ethidium bromide. These flow cytometric findings are peculiar to chromatin decondensation induced by displacement of histone H1. Conversely, chromatin decondensation caused by lowering of the divalent ion concentration, without displacement of histone H1, is characterized only by an increase in perpendicular light scatter.
Cocco
L
L
Istituto di Anatomia Umana Normale, University of Bologna, Italy.
Papa
S
S
Maraldi
N M
NM
Santi
P
P
Martelli
A M
AM
Rizzoli
R
R
Manzoli
F A
FA
D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D005434:Flow Cytometry; D006657:Histones / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D008836:Micrococcal Nuclease / Q000378:metabolism; D008854:Microscopy, Electron; D010718:Phosphatidylserines / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D002843:Chromatin; D006657:Histones; D008081:Liposomes; D010718:Phosphatidylserines; D008836:Micrococcal Nuclease