Alterations in nuclear matrix ultrastructure of G1 mammalian cells following heat shock: resinless section electron microscopy, biochemical, and immunofluorescence studies.
Heat shock is known to inhibit vital nuclear functions associated with DNA and RNA metabolism. It has been proposed that the reported heat-induced excess protein accumulation in the nuclear matrix (NM) fraction may alter NM sites crucial for DNA and RNA processing. To test this hypothesis, we examined the fine structure of the NM in synchronous populations of G1 Chinese hamster ovary cells before and after heating by using the technique of resinless section electron microscopy. Heat did induce morphological alterations in the NM. The NM of control cells contained a honeycomb-like arrangement of fibers after chromatin removal. Following heat shock, NMs appeared as more highly anastomosing networks of polymorphic fibers and an overall increase in electron density was observed. Residual nucleoli from heated NMs underwent alterations in distributions of electron density both internally and at their peripheries. The increase in electron density observed in heated NMs was accompanied by an increase in protein mass and a relatively smaller increase in RNA mass as indicated by parallel sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and isotopic labeling (protein/DNA and RNA) studies. Some excess protein accumulation could also be directly localized onto NM fibers by use of antibodies to heterogeneous ribonucleoprotein complex antigens. It is concluded that alterations of NM fine structure can reflect the heat-stressed state of the cell, may account for the heat-induced inhibition of nucleic acid metabolism, and may be useful as an indicator of physiological or pathological stress in general.
Wachsberger
P R
PR
Department of Radiation Oncology and Nuclear Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Coss
R A
RA
D000818:Animals; D016466:CHO Cells; D002458:Cell Fractionation; D002843:Chromatin / Q000648:ultrastructure; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D005455:Fluorescent Antibody Technique; D016193:G1 Phase; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006358:Hot Temperature; D015151:Immunoblotting; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D015530:Nuclear Matrix / Q000737:chemistry / Q000648:ultrastructure*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D012313:RNA / Q000032:analysis; D012261:Ribonucleoproteins / Q000032:analysis
D002843:Chromatin; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D011506:Proteins; D012261:Ribonucleoproteins; D012313:RNA; D004247:DNA