Constitutive expression of a somatic heat-inducible hsp70 gene during amphibian oogenesis

Development ◽  
1993 ◽  
Vol 119 (3) ◽  
pp. 921-932 ◽  
Author(s):  
B. Billoud ◽  
M.L. Rodriguez-Martin ◽  
L. Berard ◽  
N. Moreau ◽  
N. Angelier
Keyword(s):  
Heat Shock ◽  
Constitutive Expression ◽  
Hsp70 Gene ◽  
S1 Nuclease ◽  
Hsp70 Mrna ◽  
Protein Levels ◽  
Clear Cut ◽  
Pleurodeles Waltl ◽  
Nascent Transcripts ◽  
Related Proteins

We isolated and characterized a sequence coding for heat-shock protein 70 (HSP70) of the amphibian Pleurodeles waltl. Results from S1 nuclease protection assays led us to conclude that an hsp70 gene, strictly inducible in somatic cells during heat shock, is constitutively active during oogenesis. By quantitative northern and western blot analysis, we showed that both hsp70 mRNA and HSP70-related protein levels increased in oocytes from stage II to stage VI under physiological conditions. Furthermore, by in situ hybridization to the nascent transcripts of lampbrush chromosome loops, we provided evidence for a clear-cut relationship between this increase in hsp70 mRNA and transcriptional activity during the lampbrush stage of oogenesis. These results strongly suggest that hsp70 genes are actively transcribed throughout oogenesis. HSP70-related proteins localized in the cytoplasm of young oocytes are progressively transferred to the nucleus in the course of oogenesis and preferentially accumulated in the nuclei of some stage VI oocytes.

scholarly journals The human heat-shock protein family. Expression of a novel heat-inducible HSP70 (HSP70B′) and isolation of its cDNA and genomic DNA

1990 ◽  
Vol 267 (1) ◽  
pp. 125-132 ◽  
Author(s):  
T K Leung ◽  
M Y Rajendran ◽  
C Monfries ◽  
C Hall ◽  
L Lim
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Hela Cells ◽  
Sequence Similarity ◽  
Sequence Divergence ◽  
Hsp70 Gene ◽  
S1 Nuclease ◽  
Hsp70 Mrna ◽  
Structural And Functional Properties ◽  
Human Heat Shock Protein

The human heat-shock protein multigene family comprises several highly conserved proteins with structural and functional properties in common, but which vary in the extent of their inducibility in response to metabolic stress. We have isolated and characterized a novel human HSP70 cDNA, HSP70B′ cDNA, and its corresponding gene sequence. HSP70B′ cDNA hybrid-selected an mRNA encoding a more basic 70 kDa heat-shock protein that both the major stress-inducible HSP70 and constitutively expressed HSC70 heat-shock proteins, which in common with other heat-shock 70 kDa proteins bound ATP. The complete HSP70B′ gene was sequenced and, like the major inducible HSP70 gene, is devoid of introns. The HSP70B′ gene has 77% sequence similarity to the HSP70 gene and 70% similarity to HSC70 cDNA, with greatest sequence divergence towards the 3′-terminus. The HSP70B′ gene represents a functional gene, as indicated by Northern-blot analysis with specific oligonucleotides, hybrid-selected translation with a specific 3′ cDNA sequence and S1 nuclease protection experiments. In contrast with HSP70 mRNA, which is present at low concentrations in HeLa cells and readily induced by heat or CdCl2 treatment in both fibroblasts and HeLa cells, HSP70B′ mRNA was induced only at higher temperature and showed no basal expression. The differences in patterns of induction may be due to the special features of the promoter region of the HSP70B′ gene.

Regulation of HSP70 by PTH: a model of gene regulation not mediated by changes in cAMP levels

1996 ◽  
Vol 271 (1) ◽  
pp. C121-C129 ◽  
Author(s):  
S. Fukayama ◽  
B. Lanske ◽  
J. Guo ◽  
H. M. Kronenberg ◽  
F. R. Bringhurst
Keyword(s):  
Heat Shock ◽  
Adenylate Cyclase ◽  
Gene Transcription ◽  
Cellular Response ◽  
Host Cells ◽  
Osteoblastic Cell ◽  
Target Cells ◽  
Hsp70 Gene ◽  
Hsp70 Mrna ◽  
Human Hsp70

Parathyroid hormone (PTH) activates both adenylate cyclase and phospholipase C in target cells, and cloned PTH/PTH-related protein (PTHrP) receptor can mediate both responses when expressed in host cells such as LLC-PK1 renal epithelial cells. Because calcitonin (CT) is known to augment 70-kDa heat shock protein (HSP70) mRNA by an adenosine 3',5'-cyclic monophosphate (cAMP)-independent mechanism in LLC-PK1 cells, we examined regulation of HSP70 transcription by PTH in these cells. Like CT, human PTH-(1-34) [hPTH-(1-34); 10(-10) to 10(-7) M)] increased porcine HSP70 mRNA and human HSP70 promoter-chloramphenicol acetyltransferase (CAT) expression within 4 h in LLC-PK1 cells that stably express > or = 100,000 PTH/PTHrP receptors per cell. The effect of PTH on HSP70 mRNA was not mimicked by cAMP analogues, forskolin, phorbol esters, Ca2+ ionophores, or alpha-thrombin; was insensitive to pertussis toxin; and was not due to increased mRNA stability. The upregulation of HSP70 gene transcription by hPTH (and CT) was clearly observed even after deletion of the functional heat shock consensus element in the promoter region of the human HSP70/CAT reporter. Upregulation of HSP70 transcription via endogenous PTH receptors also was observed in the osteoblastic cell lines SaOS-2 and ROS 17/2.8. Regulation of HSP70 gene transcription by PTH may be a common cellular response to the hormone, which, in some cells, may not be mediated by activation of adenylate cyclase or protein kinase C.

scholarly journals Expression of a heat-inducible gene of the HSP70 family in human myelomonocytic cells: regulation by bacterial products and cytokines

Blood ◽  
1991 ◽  
Vol 77 (3) ◽  
pp. 579-586
Author(s):  
G Fincato ◽  
N Polentarutti ◽  
A Sica ◽  
A Mantovani ◽  
F Colotta
Keyword(s):  
Heat Shock ◽  
Transcriptional Activation ◽  
Blot Analysis ◽  
Human Peripheral Blood ◽  
Hsp70 Expression ◽  
Hsp70 Gene ◽  
Blood Leukocytes ◽  
Hsp70 Mrna ◽  
Hsp70 Family ◽  
Myelomonocytic Cells

In this study we have examined the expression of a heat-shock protein (HSP) 70 gene in normal human peripheral blood leukocytes. Northern blot analysis showed that appreciable levels of hsp70 mRNA are present in monocytes and granulocytes, whereas transcript levels were barely detectable or absent in lymphocytes. Monocytes functionally activated by bacterial lipopolysaccharide (LPS) showed an early (15 minutes) increase of hsp70 transcripts that was shown, by actinomycin D blocking and nuclear run-off experiments, to be dependent on transcriptional activation of the gene. LPS did not appreciably affect the hsp70 mRNA half-life. Monocytes exposed to inactivated streptococci, phorbol-12- myristate-13-acetate, and tumor necrosis factor showed augmented levels of hsp70 transcripts, whereas interferon-gamma and monocyte, granulocyte, and granulocyte-monocyte colony-stimulating factors had no effect. Adherence to plastic augmented hsp70 expression in monocytes. S1 protection analysis indicated that the gene expressed in monocytes is indeed a heat-inducible member of the hsp70 gene family rather than a constitutively expressed heat-shock cognate gene. Western blot analysis showed that a heat-inducible HSP72 was present in monocytes and, at augmented levels, in LPS-treated monocytes. LPS-activated monocytes were more resistant to heat shock than unstimulated cells. These data indicate that a heat-inducible hsp70 gene can be efficiently expressed in myelomonocytic cells at physiologic temperatures. Expression of hsp70 genes in monocytes suggests a possible role of heat- inducible genes in the differentiation and/or functional activation of terminally differentiated nonproliferating elements of the myelomonocytic lineage.

scholarly journals Effects of transport stress on pathological injury and expression of main heat shock proteins in the caprine stomach

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Wei Hu ◽  
Tian Ye ◽  
Yanzhen Yang ◽  
Ben Liu ◽  
Wenya Zheng
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Heat Shock Protein ◽  
Jiangxi Province ◽  
Control Group ◽  
Hsp70 Mrna ◽  
Stress Group ◽  
Protein Levels ◽  
Transport Stress ◽  
Shock Proteins

Abstract Background Transportation is necessary to introduce new breeds of goats to the farm and move the adult meat goat from the farm to the slaughterhouse. However, these actions may give rise to transport stress. Heat shock proteins (HSPs) are playing some important regulate roles during transport stress. The aim of this study was to evaluate the effects of transport stress on the pathological injury and HSPs expression in the stomach of goats. A total of three batches of Ganxi goats from western Jiangxi province were enrolled in this study. For each batch, twelve healthy adult male goats were randomly divided into three groups (four goats per batch and per group): Control group, stress group transported during 2 h and stress group transported during 6 h. Results Our results showed that the different degrees of stomach walls damage, with the change of expression levels of heat shock protein 27 (HSP27), heat shock protein 70 (HSP70) and heat shock protein 90 (HSP90), occurred after goats transportation. In rumen, the mRNA and protein expressions of HSP27 and HSP70 were increased after transport stress, but not HSP90. In reticulum, all three HSPs mRNA and protein levels were upregulated after 2 h transport, but decreased after 6 h transport. In omasum, HSP27 and HSP70 mRNA and protein were increased after transport stress, however, HSP90 mRNA level only had a slightly enhancement after transport stress. In abomasum, HSP70 and HSP90 mRNA and protein levels were increased after transport stress, but HSP27 was decreased after transport stress. Conclusions Taken together, these results revealed that the pathological changes in the gastric tissues and the stomach HSPs expression in goats are related to transport stress and duration. Moreover, this study also provides some new data to advocate reducing transport stress of goats and improving animal welfare.

Developmental-stage-specific expression of the hsp70 gene family during differentiation of the mammalian male germ line

1987 ◽  
Vol 7 (5) ◽  
pp. 1791-1796
Author(s):  
Z F Zakeri ◽  
D J Wolgemuth
Keyword(s):  
Heat Shock ◽  
Gene Family ◽  
Germ Line ◽  
Cell Lineage ◽  
Hsp70 Gene ◽  
Developmentally Regulated ◽  
Specific Expression ◽  
Hsp70 Mrna ◽  
Somatic Tissues ◽  
Exogenous Stress

Mouse somatic tissues contain low levels of transcripts homologous to the heat shock-inducible and cognate members of the heat shock protein 70 (hsp70) gene family. An abundant, unique sized hsp70 mRNA of 2.7 kilobases (kb) is present in testes in the absence of exogenous stress. Its expression is restricted to germ cells and is developmentally regulated. The 2.7-kb transcript first appears during the haploid phase of spermatogenesis and is stable throughout the morphogenic stages of spermiogenesis. A 2.7-kb hsp70 mRNA is present in rat and human testes. These observations suggest that a member of the hsp70 gene family plays a role in the development of the mammalian male germ cell lineage.

Heat shock-induced translational control of HSP70 and globin synthesis in chicken reticulocytes

1984 ◽  
Vol 4 (11) ◽  
pp. 2437-2448
Author(s):  
S S Banerji ◽  
N G Theodorakis ◽  
R I Morimoto
Keyword(s):  
Heat Shock ◽  
Translational Control ◽  
Actinomycin D ◽  
Elevated Temperatures ◽  
Rapid Change ◽  
Hsp70 Gene ◽  
Globin Mrna ◽  
Hsp70 Mrna ◽  
Globin Synthesis

Incubation of chicken reticulocytes at elevated temperatures (43 to 45 degrees C) resulted in a rapid change in the pattern of protein synthesis, characterized by the decreased synthesis of normal proteins, e.g., alpha and beta globin, and the preferential and increased synthesis of only one heat shock protein, HSP70. The repression of globin synthesis was not due to modifications of globin mRNA because the level of globin mRNA and its ability to be translated in vitro were unaffected. The HSP70 gene in reticulocytes was transcribed in non-heat-shocked cells, yet HSP70 was not efficiently translated until the cells had been heat shocked. In non-heat-shocked reticulocytes, HSP70 mRNA was a moderately abundant mRNA present at 1 to 2% of the level of globin mRNA. The rapid 20-fold increase in the synthesis of HSP70 after heat shock was not accompanied by a corresponding increase in the rate of transcription of the HSP70 gene or accumulation of HSP70 mRNA. These results suggest that the elevated synthesis of HSP70 is due to the preferential utilization of HSP70 mRNA in the heat-shocked reticulocyte. The heat shock-induced alterations in the reticulocyte protein-synthetic apparatus were not reversible. Upon return to control temperatures (37 degrees C), heat-shocked reticulocytes continued to synthesize HSP70 at elevated levels whereas globin synthesis continued to be repressed. Despite the presence of HSP70 mRNA in non-heat-shocked reticulocytes, we found that continued transcription was necessary for the preferential translation of HSP70 in heat-shocked cells. Preincubation of reticulocytes with the transcription inhibitor actinomycin D or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole blocked the heat shock-induced synthesis of HSP70. Because the level of HSP70 mRNA was only slightly diminished in cells treated with actinomycin D, we suggest two possible mechanisms for the preferential translation of HSP70 mRNA: the translation of only newly transcribed HSP70 mRNA or the requirement of a newly transcribed RNA-containing factor.

Constitutive roles for inducible genes: evidence for the alteration in expression of the inducible hsp70 gene in Antarctic notothenioid fishes

2004 ◽  
Vol 287 (2) ◽  
pp. R429-R436 ◽  
Author(s):  
Sean P. Place ◽  
Mackenzie L. Zippay ◽  
Gretchen E. Hofmann
Keyword(s):  
New Zealand ◽  
Heat Shock ◽  
Thermal Stresses ◽  
Expression Patterns ◽  
Antarctic Fish ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
Hsp70 Mrna ◽  
Inducible Genes ◽  
The Antarctic

Previous research on the Antarctic notothenioid fish Trematomus bernacchii demonstrated the loss of the heat shock response (HSR), a classical cellular defense mechanism against thermal stress, characterized by the rapid synthesis of heat shock proteins (Hsps). In the current study, we examined potential mechanisms for the apparent loss of the HSR in Antarctic notothenioids and, in addition, compared expression patterns of two genes from the 70-kDa Hsp family ( hsc71 and hsp70) in tissues from T. bernacchii to expression patterns in tissues of two closely related temperate notothenioid fishes from New Zealand, Bovichtus variegatus and Notothenia angustata. The results showed that transcript for both the constitutive and inducible genes in the Hsp70 gene family were expressed in detectable levels in all three species. However, only the cold-temperate New Zealand fishes displayed the ability to upregulate the inducible transcript, hsp70. Although hsp70 was present in detectable levels in several tissues of the Antarctic notothen T. bernacchii, in vitro thermal stresses failed to produce a significant increase in mRNA levels. In all species, the expression of the constitutive transcript hsc71 was variable and nonresponsive to temperature increases, even at temperatures as high as 10°C above the ecologically relevant range for the species under study. Field-collected tissues from T. bernacchii (sampled immediately after capture) indicated that hsp70 mRNA was expressed at high levels in field-acclimatized fishes. Thus upregulation of molecular chaperones suggested that low-temperature stress may be significantly denaturing to cellular proteins in Antarctic fish, an observation that was supported by elevated levels of ubiquitin-conjugated protein.

scholarly journals Structure and expression of the human gene encoding major heat shock protein HSP70.

1985 ◽  
Vol 5 (2) ◽  
pp. 330-341 ◽  
Author(s):  
B Wu ◽  
C Hunt ◽  
R Morimoto
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Genomic Library ◽  
Human Gene ◽  
Chimeric Gene ◽  
Hsp70 Gene ◽  
Gene Encoding ◽  
Hsp70 Mrna ◽  
Flanking Sequences ◽  
Heat Shock Protein Hsp70

We have cloned a human gene encoding the 70,000-dalton heat shock protein (HSP70) from a human genomic library, using the Drosophila HSP70 gene as a heterologous hybridization probe. The human recombinant clone hybridized to a 2.6-kilobase polyadenylated mRNA from HeLa cells exposed to 43 degrees C for 2 h. The 2.6-kilobase mRNA was shown to direct the translation in vitro of a 70,000-dalton protein similar in electrophoretic mobility to the HSP70 synthesized in vivo. From the analysis of S1 nuclease-resistant mRNA-DNA hybrids, the HSP70 gene appears to be transcribed as an uninterrupted mRNA of 2.3 kilobases. We show that the cloned HSP70 gene contains the sequences necessary for heat shock-induced expression by two criteria. First, hamster cells transfected with a subclone containing the HSP70 gene and flanking sequences synthesized a HSP70-like protein upon heat shock. Second, human cells transfected with a chimeric gene containing the 5' flanking sequences of the HSP70 gene and the coding sequences of the bacterial chloramphenicol acetyltransferase gene transcribed the chimeric gene upon heat shock. We show that the HSP70 mRNA transcribed in an adenovirus 5 transformed human cell line (293 cells) is identical to the HSP70 mRNA induced by heat shock.

scholarly journals Induction of HSP70 gene expression by the antiproliferative prostaglandin PGA2: a growth-dependent response mediated by activation of heat shock transcription factor.

1992 ◽  
Vol 12 (4) ◽  
pp. 1528-1534 ◽  
Author(s):  
N J Holbrook ◽  
S G Carlson ◽  
A M Choi ◽  
J Fargnoli
Keyword(s):  
Heat Shock ◽  
Stress Proteins ◽  
Heat Shock Factor ◽  
Hsp70 Expression ◽  
Hsp70 Gene ◽  
Hsp70 Mrna ◽  
Antiproliferative Effects ◽  
Growth State ◽  
Hsp70 Family ◽  
High Level

Prostaglandins (PG) of the A series are potent inhibitors of cell proliferation. Recently, it was shown that PGA2-induced growth arrest was associated with the increased synthesis of stress proteins encoded by the HSP70 gene family. In this study, we have examined the molecular basis for this increases HSP70 expression. Northern (RNA) blot analysis and nuclear run-on assays demonstrated that induction of high levels of HSP70 mRNA results from an increase in the rate of transcription. High-level induction is specific to the HSP70 family of heat shock proteins and is rapid, reversible, dose dependent, and specific for PGs capable of growth-arresting HeLa cells. In addition, the response was found to be highly dependent on the growth state of the cells, as induction occurs in growing but not in confluent nongrowing cell populations. Induction is dependent on the activation of heat shock factor. Cycloheximide pretreatment, which inhibits the antiproliferative effects of PGA2, prevents activation of the heat shock factor and induction of HSP70 mRNA by PGA2. These results support a role for HSP70 in mediating the antiproliferative effects of PGA2.

Schistosome extracts with heat shock factor activity revealed by the gel shift assay

Parasitology ◽  
1994 ◽  
Vol 108 (1) ◽  
pp. 35-42 ◽  
Author(s):  
R. Levy-Holtzman ◽  
I. Schechter
Keyword(s):  
Heat Shock ◽  
Transcriptional Activation ◽  
Developmental Stages ◽  
Consensus Sequence ◽  
Structural Features ◽  
Heat Shock Factor ◽  
Regulatory Function ◽  
Hsp70 Gene ◽  
Mobility Shift ◽  
Hsp70 Mrna

SUMMARYTo understand the regulated expression of stage-specific genes of schistosomes, it is necessary to identify regulatory DNA elements and DNA-binding proteins that control the level of gene transcription. Here we describe the preparation of Schistosoma mansoni extracts with active transcription factors detected by the electrophoretic mobility shift assay (EMSA). We analysed the hsp70 system of S. mansoni because the promoters of the hsp70 gene contain two heat shock elements (HSE) that differ from the consensus sequence (CnnGAAnnTTCnnG) at one (HSEI) or three (HSEII) positions, and it is known that transcriptional activation of hsp70 genes is mediated by interaction of HSE with the heat shock factor (HSF). Analyses of parasite extracts from different developmental stages demonstrate the presence of putative HSF that correlates with the pattern of hsp70 mRNA expression (cercariae-, schistosomula+, adult worms+). Cercarial extracts did not show binding of 32P-labelled HSEI or HSEII. Extracts of schistosomula and of adult worms kept at 37 or 42 °C showed binding of HSEI but not HSEII. The specificity of HSEI-HSF complex formation was ascertained by inhibition experiments. The EMSA experiments and structural features of the hsp70 promoter indicate that HSEI is the major DNA element responsible for transcriptional activation of the hsp70 gene, while the HSEII may be a redundant sequence with minor (if any) regulatory function. The extracts reported here can be used to study transcriptional control of other schistosome genes.

Sign in / Sign up

Export Citation Format

Share Document