Specific intranucleolar distribution of Hsp70 during heat shock in polytene cells.
Hsp70, the most abundant and conserved heat shock protein, has been described as strongly concentrating in the nucleolus during heat shock. The important metabolic processes that take place in the nucleolus, rDNA transcription, processing, and assembling with ribosomal proteins, and the nucleolar architecture itself are very sensitive to temperature changes. In this work, we have analyzed in detail the nucleolar changes, in structure and activity, induced by temperature in Chironomus thummi salivary gland cells and the fine subnucleolar localization of Hsp70 during heat shock. The optimum temperature chosen to induce the heat shock response was 35 degrees C. Under these conditions transcription of heat shock genes, inactivation of previously active genes and maximum synthesis of Hsps take place, while survival of larvae and recovery were ensured. After 1 h at 35 degrees C, nucleoli change from a uniform control pattern to a segregated pattern of nucleolar components that can be observed even at the light microscopic level. The dense fibrillar component (DFC) and the granular component appeared perfectly differentiated and spatially separated, the former occupying mainly the central inner region surrounded by a rim of granular component. Hsp70 was specifically localized within the DFC upon heat shock as shown by immunolocalization by both light and electron microscopy. Pulse labeling with [3H]uridine proves that rRNA transcription continues during heat shock. The pattern of Hsp70 distribution within the nucleolus correlates with that of newly produced rRNA transcripts. Hsp70 also colocalizes with RNA polymerase I, both being restricted to the DFC. These data show that the DFC seems to be the intranucleolar target for Hsp70 in heat-shocked cells. We discuss these results in relation to the possible function of Hsp70 in the first steps of preribosome synthesis.
Morcillo
G
G
Department of Organic Chemistry and Biology, Universidad Nacional de Educación a Distancia, Madrid, Spain. gmorcill@sr.uned.es
Gorab
E
E
Tanguay
R M
RM
Díez
J L
JL
D000818:Animals; D002451:Cell Compartmentation; D002466:Cell Nucleolus / Q000648:ultrastructure*; D002683:Chironomidae; D002875:Chromosomes / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology / Q000302:isolation & purification*; D018869:Heat-Shock Response; D007814:Larva; D016253:Microscopy, Immunoelectron; D012318:RNA Polymerase I / Q000276:immunology / Q000302:isolation & purification; D012322:RNA Precursors / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000302:isolation & purification; D012469:Salivary Glands
D018840:HSP70 Heat-Shock Proteins; D012322:RNA Precursors; D012335:RNA, Ribosomal; D012318:RNA Polymerase I