Dynamic changes in intracellular localization and isoforms of the 27-kD stress protein in human keratinocytes

We have begun to characterize the low molecular weight, 27-kD heat shock or stress protein (HSP27) in normal keratinocytes and in HaCaT, a spontaneously transformed keratinocyte line. The presence and location of HSP27 was determined by indirect immunofluorescence on fixed whole cells and immunoblot analysis of cytosolic, membrane, nuclear, and cytoskeletal cell fractions. HSP27 is localized throughout the cytoplasm of cells at 37 degrees C. After heating at 42 degrees C, there is a rapid (within 10 min) increase in nuclear HSP27. Two-dimensional gel analysis of whole cell HaCaT lysates identified multiple isoforms of HSP27 with different isoelectric points. The function of HSP27 is largely unknown but its presence throughout the cytoplasm of cells at 37 degrees C, its translocation to the nucleus after cellular stress, and the presence of multiple isoforms suggest a biologic role in both stressed and unstressed human keratinocytes.