Induction and intracellular localization of the 80-kilodalton heat-shock protein of Neurospora crassa

1992 ◽  
Vol 70 (12) ◽  
pp. 1347-1355 ◽  
Author(s):  
H. S. Roychowdhury ◽  
T. J. MacAlister ◽  
J. W. Costerton ◽  
M. Kapoor
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Neurospora Crassa ◽  
Immunoelectron Microscopy ◽  
Intracellular Localization ◽  
Normal Growth ◽  
Immunogold Labelling ◽  
Subcellular Compartment ◽  
Intracellular Location ◽  
Gold Label

The most abundant heat-shock protein of Neurospora crassa is a multimeric glycoprotein of 80-kilodaltons (i.e., HSP80), induced strongly by hyperthermia and at a lower level by sodium arsenite, ethanol, and carbon source depletion. Immunoelectron microscopy, using indirect immunogold labelling demonstrated that HSP80 was undetectable in mycelium cultured at the normal growth temperature of 28 °C, but it appeared rapidly following the commencement of heat-shock treatment at 48 °C. HSP80, visualized by the gold label, was observed almost exclusively in the cytoplasm, exhibiting a uniform distribution. Association of this protein with cellular membranes and (or) targeting to a particular subcellular compartment or organelle was not apparent.Key words: 80-kilodalton heat-shock protein, Neurospora, intracellular location, immunoelectron microscopy.

scholarly journals The intracellular location of yeast heat-shock protein 26 varies with metabolism.

1989 ◽  
Vol 108 (2) ◽  
pp. 425-439 ◽  
Author(s):  
J M Rossi ◽  
S Lindquist
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Physiological State ◽  
Intracellular Localization ◽  
Normal Growth ◽  
Genetically Engineered ◽  
Wild Type ◽  
Intracellular Location ◽  
Small Hsps ◽  
In Cells

An antibody highly specific for heat-shock protein (hsp)26, the unique small hsp of yeast, and mutants carrying a deletion of the HSP26 gene were used to examine the physical properties of the protein and to determine its intracellular distribution. The protein was found in complexes with a molecular mass of greater than 500 kD. Thus, it has all of the characteristics, including sequence homology and induction patterns, of small hsps from other organisms. When log-phase cells growing in glucose were heat shocked, hsp26 concentrated in nuclei and continued to concentrate in nuclei when these cells were returned to normal temperatures for recovery. However, hsp26 did not concentrate in nuclei under a variety of other conditions. For example, in early stationary-phase cells hsp26 is induced at normal growth temperatures. This protein was generally distributed throughout the cells, even after heat shock. Similarly, in cells genetically engineered to synthesize hsp26 in the presence of galactose, hsp26 did not concentrate in nuclei, with or without a heat shock. To determine if the failure of hsp26 to concentrate in the nucleus of these cells was due to the fact that the protein had been produced at 25 degrees C or to a difference in the physiological state of the cell, we investigated the distribution of the heat-induced protein in cells grown under several different conditions. In wild-type cells grown in galactose or acetate and in mitochondrial mutants grown in glucose or galactose, hsp26 also failed to concentrate in nuclei with a heat shock. We conclude that the intracellular location of hsp26 in yeast depends upon the physiological state of the cell and not simply upon the presence or absence of heat stress. Our findings may explain why previous investigations of the intracellular localization of small hsps in a variety of organisms have yielded seemingly contradictory results.

Ethanol and carbon-source starvation enhance the accumulation of HSP80 in Neurospora crassa

1988 ◽  
Vol 34 (2) ◽  
pp. 162-168 ◽  
Author(s):  
H. S. Roychowdhury ◽  
M. Kapoor
Keyword(s):  
Heat Shock ◽  
Neurospora Crassa ◽  
Carbon Catabolite Repression ◽  
Normal Growth ◽  
Carbon Starvation ◽  
High Molecular Mass ◽  
Sucrose Starvation ◽  
Shock Response ◽  
Shock Proteins ◽  
Starvation Conditions

In Neurospora crassa, heat shock results in the induction of 9 to 11 heat shock proteins (HSP), of which HSP80 is the most abundant and the first to be synthesized. The induction of HSP80 was investigated during normal growth (2% sucrose) and under sucrose starvation. Transfer of mycelium to a medium supplemented with ethanol stimulated the synthesis of HSP80, even at the normal growth temperature of 28 °C. It was also synthesized under carbon starvation conditions, where the medium was supplemented with 0.02% sucrose, 0.3% acetate, 0.2% lactate, or ethanol. A 30–35 kilodalton polypeptide was induced by heat shock in carbon-sufficient media, but in 0.02% sucrose and 0.3% acetate containing media it was synthesized at normal temperatures. While the overall heat shock response remained unaltered in these cultures, the abundance of HSP90 and HSP70, relative to HSP80, was greater. HSP80 appears to be controlled by carbon-catabolite repression as well as heat shock. Another high molecular mass protein (tentatively designated alc'80') was observed to be induced by heat shock, provided carbon starvation conditions prevailed concurrently.

scholarly journals Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein.

1989 ◽  
Vol 9 (6) ◽  
pp. 2615-2626 ◽  
Author(s):  
E Hickey ◽  
S E Brandon ◽  
G Smale ◽  
D Lloyd ◽  
L A Weber
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Normal Growth ◽  
Gene Families ◽  
Growth Conditions ◽  
Complete Sequence ◽  
Start Codon ◽  
Hybridization Probe ◽  
Reading Frame ◽  
Alpha Gene

Vertebrate cells synthesize two forms of the 82- to 90-kilodalton heat shock protein that are encoded by distinct gene families. In HeLa cells, both proteins (hsp89 alpha and hsp89 beta) are abundant under normal growth conditions and are synthesized at increased rates in response to heat stress. Only the larger form, hsp89 alpha, is induced by the adenovirus E1A gene product (M. C. Simon, K. Kitchener, H. T. Kao, E. Hickey, L. Weber, R. Voellmy, N. Heintz, and J. R. Nevins, Mol. Cell. Biol. 7:2884-2890, 1987). We have isolated a human hsp89 alpha gene that shows complete sequence identity with heat- and E1A-inducible cDNA used as a hybridization probe. The 5'-flanking region contained overlapping and inverted consensus heat shock control elements that can confer heat-inducible expression on a beta-globin reporter gene. The gene contained 10 intervening sequences. The first intron was located adjacent to the translation start codon, an arrangement also found in the Drosophila hsp82 gene. The spliced mRNA sequence contained a single open reading frame encoding an 84,564-dalton polypeptide showing high homology with the hsp82 to hsp90 proteins of other organisms. The deduced hsp89 alpha protein sequence differed from the human hsp89 beta sequence reported elsewhere (N. F. Rebbe, J. Ware, R. M. Bertina, P. Modrich, and D. W. Stafford (Gene 53:235-245, 1987) in at least 99 out of the 732 amino acids. Transcription of the hsp89 alpha gene was induced by serum during normal cell growth, but expression did not appear to be restricted to a particular stage of the cell cycle. hsp89 alpha mRNA was considerably more stable than the mRNA encoding hsp70, which can account for the higher constitutive rate of hsp89 synthesis in unstressed cells.

scholarly journals Human cyclophilin 40 is a heat shock protein that exhibits altered intracellular localization following heat shock

2001 ◽  
Vol 6 (1) ◽  
pp. 59 ◽  
Author(s):  
Peter J. Mark ◽  
Bryan K. Ward ◽  
Premlata Kumar ◽  
Hooshang Lahooti ◽  
Rodney F. Minchin ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Intracellular Localization ◽  
Cyclophilin 40

Heat shock response in Neurospora crassa: purification and some properties of HSP 80

1990 ◽  
Vol 68 (10) ◽  
pp. 1218-1221 ◽  
Author(s):  
H. S. Roychowdhury ◽  
M. Kapoor
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Neurospora Crassa ◽  
Heat Shock Response ◽  
Periodic Acid ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Molecular Size ◽  
Periodic Acid Schiff ◽  
Shock Response

The heat shock response of Neurospora crassa was investigated. A 80-kilodalton heat shock protein (HSP 80) was purified to near homogeneity from heat-shocked mycelial extracts employing ammonium sulphate fractionation, gel filtration, and ion-exchange and affinity chromatography. It was observed to migrate as a single band on one-dimensional sodium dodecyl sulphate – polyacrylamide gels, with a molecular mass of ~ 83 kilodaltons (kDa). On two-dimensional gels it resolved into four polypeptide species with isoelectric points in the acidic range, which on staining with periodic acid – Schiff method were demonstrated to be glycosylated. In the native state, HSP 80 had a molecular size of ~610 kDa.Key words: Neurospora, heat shock, fast protein liquid chromatography, 80-kilodalton heat shock protein, glycoprotein.

Interaction of Drosophila 27,000 Mr heat-shock protein with the nucleus of heat-shocked and ecdysone-stimulated culture cells

1989 ◽  
Vol 92 (1) ◽  
pp. 29-36 ◽  
Author(s):  
J.F. Beaulieu ◽  
A.P. Arrigo ◽  
R.M. Tanguay
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Elevated Temperatures ◽  
Immunofluorescence Microscopy ◽  
Intracellular Localization ◽  
Molting Hormone ◽  
Culture Cells ◽  
Ionic Detergent ◽  
Detergent Extraction ◽  
Nuclear Component

The intracellular localization and expression of hsp27 (heat-shock protein 27) were investigated by cellular fractionation and immunofluorescence microscopy in Drosophila S3 cells. In unstressed cells, hsp27 is expressed in only 2% of the cells, whereas following heat shock, during recovery or after induction by ecdysone, the protein is detected in all cells. Under all these conditions, hsp27 appears to be concentrated in the nuclear region as revealed by immunofluorescence. During heat shock, this hsp is localized primarily in the nucleus with an enrichment in the perinucleolar region. However, the cellular fractionation data indicate that the nature of hsp27 interaction with nuclear components greatly differs depending on whether or not cells were subjected to elevated temperatures. After heat shock, hsp27 is resistant to non-ionic detergent extraction. In cells allowed to recover at normal temperature and in those where its synthesis was induced by the molting hormone, ecdysone, this hsp is readily solubilized by detergent. These data suggest that, following heat shock, hsp27 may become physically associated with some nuclear component(s) that are resistant to detergent extraction.

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