Distribution of nuclear matrix proteins in interphase CHO cells and rearrangements during the cell cycle. An ultrastructural study.
The nuclear matrix contains a group of residual non-histone proteins which remain structurally organized after extensive extraction of isolated nuclei with a high salt buffer, nucleases and a non-ionic detergent. Electron microscopic examination shows that the nuclear matrix is composed of a pore-complex lamina, an intranuclear network and residual nucleoli. In CHO cells biochemical analyses performed by one-dimensional SDS-PAGE show three major nuclear matrix polypeptides with molecular weights between 60 and 70 kDa. Polyclonal antibodies produced against these polypeptides were used to determine their nuclear distribution. Using immunoblotting, these proteins were found in whole nuclei, nuclear matrix, and in the intranuclear network but not in the pore-complex lamina. In order to determine the relationship between these structural proteins and the organization of the nucleus, the proteins were localized in situ. Ultrastructural detection was carried out by immunogold staining of thin sections of Lowicryl K4M-embedded cells. In interphase nuclei all condensed chromatin clumps were labelled. The nucleolus and the interchromatin granules were never immunogold-stained. During mitosis, the label was found to be associated with the chromosomes. This study shows that unlike the lamins, these 60-70 kDa nuclear matrix proteins are associated with the condensed chromatin throughout the cell cycle.
Noaillac-Depeyre
J
J
Laboratoire de Biologie Cellulaire, Centre de Biochimie et Génétique Cellulaires du CNRS, Toulouse, France.
Azum
M C
MC
Geraud
M L
ML
Mathieu
C
C
Gas
N
N
D000818:Animals; D002467:Cell Nucleus / Q000502:physiology* / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000276:immunology / Q000502:physiology*; D005260:Female; D006046:Gold; D006168:Guinea Pigs; D007106:Immune Sera; D007150:Immunohistochemistry; D007399:Interphase; D008854:Microscopy, Electron; D008938:Mitosis; D008970:Molecular Weight; D010053:Ovary / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure
D002868:Chromosomal Proteins, Non-Histone; D007106:Immune Sera; D006046:Gold