Further considerations on the thermal stabilization of the nuclear matrix in mouse erythroleukemia cells.
The morphology and the polypeptide composition of the nuclear matrix obtained from 37 degrees C incubated nuclei has been studied in mouse erythroleukemia cells. From a structural point of view, in the absence of heat treatment, the matrix lacked identifiable nucleolar remnants and the internal fibrogranular meshwork whereas a peripheral lamina was seen. On the contrary, the matrix obtained from heat exposed nuclei displayed very electrondense nucleolar remnants and an abundant inner network. These results were obtained irrespective of the type of extracting agent (2M NaCl or 0.2 M (NH4)2SO4) used to remove histones and other soluble proteins. The heat stabilization of the matrix could not be prevented by sulfhydryl blocking chemicals such as iodoacetamide and n-ethylmaleimide, thus suggesting that heat does not stabilize the matrix by inducing the formation of disulfide bonds. Only limited differences in the polypeptide pattern of matrix isolated under different conditions were seen using one-dimensional pore gradient polyacrylamide gels stained with both Coomassie Brilliant Blue and silver despite the fact that the matrix fraction from heat treated nuclei retained about three fold more protein in comparison with controls. The same results were obtained also by means of two-dimensional non-equilibrium gel electrophoresis.
Martelli
A M
AM
Istituto di Anatomia Umana Normale, Bologna, Italy.
Falcieri
E
E
Gobbi
P
P
Manzoli
L
L
Cataldi
A
A
Rana
R A
RA
Cocco
L
L
D000645:Ammonium Sulfate; D000818:Animals; D002458:Cell Fractionation; D002466:Cell Nucleolus / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005622:Friend murine leukemia virus; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D008854:Microscopy, Electron; D015530:Nuclear Matrix / Q000737:chemistry / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D012965:Sodium Chloride; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature; D013776:Tetrathionic Acid; D014407:Tumor Cells, Cultured
D009687:Nuclear Proteins; D010455:Peptides; D013439:Sulfhydryl Reagents; D012965:Sodium Chloride; D013776:Tetrathionic Acid; D000645:Ammonium Sulfate