Subcellular localization of the 84,000 dalton heat-shock protein in mouse neuroblastoma cells: evidence for a cytoplasmic and nuclear location.
Using affinity-purified antibodies, the 84,000 dalton heat-shock protein (hsp) has been localized in mouse N2A neuroblastoma cells by immunocytochemical techniques. Immunofluorescence microscopy showed that hsp84 was present both in the cytoplasm and in the nucleus. The nucleoli were found to be unlabelled. Immunogold labelling on ultrathin cryosections revealed that hsp84 was evenly distributed throughout the entire cytoplasm. No preferential association of hsp84 with the plasma membrane or with membranes from organelles was observed. In the nucleus the hsp84 was present in both the euchromatin and heterochromatin. In the nucleolus only the fibrillar part was labelled and virtually no gold particles were observed in the granular part. A long-term hyperthermic treatment of 3 h at 42.5 degrees C was found to induce an accumulation of hsp84 inside the nucleus. No alterations in hsp84 distribution were observed during a treatment of the cells with 75 microM sodium arsenite for 3 h. Drastic alterations were observed in the nucleoli after both stress treatments. The granular part had totally disappeared and only remnants of the fibrillar part which contained hsp84, were found. Besides the nuclear accumulations of hsp84 during heat shock, no additional changes in the hsp84 location in stressed cells were observed. During a recovery from the heat shock by replacing the cells at 37 degrees C, a decrease in the nuclear location of hsp84 was observed, indicating the reversibility of this process. The significance of these results for the role of hsp84 in normal and in stressed cells is discussed.
van Bergen en Henegouwen
P M
PM
Berbers
G
G
Linnemans
W A
WA
van Wijk
R
R
D000818:Animals; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D006360:Heat-Shock Proteins / Q000032:analysis*; D006358:Hot Temperature; D051379:Mice; D008854:Microscopy, Electron; D008970:Molecular Weight; D009447:Neuroblastoma / Q000473:pathology / Q000648:ultrastructure*
D006360:Heat-Shock Proteins