title: Effect of ATP on the release of hsp 70 and hsp 40 from the nucleus in heat-shocked HeLa cells.
abstract: We have recently found a novel 40-kDa heat-shock protein (hsp 40) in mammalian and avian cells and reported that the N-terminal amino acid sequence of mammalian hsp 40 has homology with the bacterial DnaJ heat-shock protein. Also, hsp 40 has been shown to be translocated from the cytoplasm into the nuclei/nucleoli by heat shock and colocalized with hsc 70 (p73) in the nucleoli of exactly the same cells. We here investigated the effect of ATP on the release of hsp 70 (both constitutive p73 and inducible p72) and hsp 40 from the nuclei/nucleoli of heat-shocked HeLa cells which were permeabilized with Nonidet-P40 using immunofluorescence and immunoblotting. Hsp 70 in the nucleoli was released by the addition of ATP but not by ADP, GTP, nonhydrolyzable ATP, nor high salt buffer. In contrast, hsp 40 was not released from the nucleoli with any of these treatments or any combination of these treatments. Thus, hsp 40 might dissociate spontaneously from the nucleoli after hsp 70 has been released in an ATP-dependent manner. Using cell fractionation methods, we showed that while the majority of hsp 40 is localized in the cytoplasm, a small portion of it is located in the microsome fraction in non-heat-shocked control cells and in cells which recovered from heat shock.
authors.affiliation: Laboratory of Experimental Radiology, Aichi Cancer Center Research Institute, Nagoya, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohtsuka
authors.forename: K R
authors.initials: KR
authors.lastname: Utsumi
authors.forename: T
authors.initials: T
authors.lastname: Kaneda
authors.forename: H
authors.initials: H
authors.lastname: Hattori
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D015153:Blotting, Western; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D066298:In Vitro Techniques; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; D000255:Adenosine Triphosphate