Rapid effect of heat shock on two heterogeneous nuclear ribonucleoprotein-associated antigens in HeLa cells.
During severe heat shock, which known to interrupt both splicing of RNA transcripts and nucleocytoplasmic transport, it is to be expected that the substructure of heterogeneous nuclear ribonucleoproteins (hnRNP) is altered in some way. Recently, we have shown that such a stress actually induces rapid alterations at the level of individual proteins (Lutz, Y., M. Jacob, and J.-P. Fuchs. 1988 Exp. Cell Res. 175:109-124). Here we report further investigations on two related 72.5-74-kD hnRNP proteins whose behavior is also rapidly modified by a heat shock at 45 degrees C, whereas no effect is observed at 42 degrees C. Using a monoclonal antibody, we show that in situ the antigens are available only when the cells are heat shocked at 45 degrees C. Subcellular fractionation shows that in normal cells the antigens are associated with the bulk of hnRNP (50-200S). During heat shock, whereas the overall characteristics of the bulk of preexisting hnRNP are unchanged, these antigens rapidly switch to a subpopulation of hnRNP with larger average size (50 to less than 300S) and increased stability. Structural analysis of the associated hnRNP in normal and stressed cells shows that in both cases the antigens are associated with the nuclear matrix subcomplex of hnRNP, which in situ is part of the internal nuclear matrix. Such hnRNP antigens, which are rapidly redistributed during a heat shock at the upper temperature range of the stress response, might well be involved in splicing and/or transport control.
Mähl
P
P
Laboratoire Génétique Moléculaire des Eucaryotes du Centre National de la Recherche Scientifique, Strasbourg, France.
Lutz
Y
Y
Puvion
E
E
Fuchs
J P
JP
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens / Q000032:analysis; D015153:Blotting, Western; D002467:Cell Nucleus / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells / Q000378:metabolism / Q000648:ultrastructure; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006358:Hot Temperature; D006801:Humans; D007525:Isoelectric Focusing; D008854:Microscopy, Electron; D008970:Molecular Weight; D010449:Peptide Mapping; D012332:RNA, Heterogeneous Nuclear / Q000096:biosynthesis*; D012261:Ribonucleoproteins / Q000096:biosynthesis* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens; D006360:Heat-Shock Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D012332:RNA, Heterogeneous Nuclear; D012261:Ribonucleoproteins