scholarly journals Cell cycle control of the human HSP70 gene: implications for the role of a cellular E1A-like function.

1985 ◽  
Vol 5 (4) ◽  
pp. 628-633 ◽  
Author(s):  
H T Kao ◽  
O Capasso ◽  
N Heintz ◽  
J R Nevins
Keyword(s):  
Cell Cycle ◽  
Heat Shock ◽  
Cell Cycle Control ◽  
Peak Level ◽  
Fold Increase ◽  
Hsp70 Expression ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
Cellular Activity ◽  
E1a Gene

The gene encoding the human 70-kilodalton heat shock protein (HSP70) is subject to activation by the adenovirus E1A gene product and appears to be regulated in the absence of heat shock by a cellular activity similar to E1A. Given the relation of E1A to alteration of growth control, we have investigated the expression of the HSP70 gene during the cell cycle. Assay of mRNA levels after release from a thymidine-aphidicolin block revealed a 20-fold increase in mRNA abundance, reaching a peak level in the post-S-phase period. Upon reaching this peak level, the abundance of the mRNA then declined as the cells entered the next cycle. Control of the abundance of the mRNA during the cell cycle appeared to be primarily at the level of transcription as measured in nuclear runoff assays. Very similar results were obtained by analyzing the expression of the HSP70 gene in the adenovirus-transformed 293 cell line. Furthermore, the E1A gene was also found to be cell cycle regulated; the activation and peak level of the E1A mRNA occurred at an earlier time than those of the heat shock mRNA, consistent with, but not proof of, the hypothesis that E1A is responsible for the cell cycle control of the HSP70 expression. We therefore suggest that the E1A-like cellular activity may govern certain aspects of cell cycle transcription.

Cell cycle control of the human HSP70 gene: implications for the role of a cellular E1A-like function

1985 ◽  
Vol 5 (4) ◽  
pp. 628-633
Author(s):  
H T Kao ◽  
O Capasso ◽  
N Heintz ◽  
J R Nevins
Keyword(s):  
Cell Cycle ◽  
Heat Shock ◽  
Cell Cycle Control ◽  
Peak Level ◽  
Fold Increase ◽  
Hsp70 Expression ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
Cellular Activity ◽  
E1a Gene

The gene encoding the human 70-kilodalton heat shock protein (HSP70) is subject to activation by the adenovirus E1A gene product and appears to be regulated in the absence of heat shock by a cellular activity similar to E1A. Given the relation of E1A to alteration of growth control, we have investigated the expression of the HSP70 gene during the cell cycle. Assay of mRNA levels after release from a thymidine-aphidicolin block revealed a 20-fold increase in mRNA abundance, reaching a peak level in the post-S-phase period. Upon reaching this peak level, the abundance of the mRNA then declined as the cells entered the next cycle. Control of the abundance of the mRNA during the cell cycle appeared to be primarily at the level of transcription as measured in nuclear runoff assays. Very similar results were obtained by analyzing the expression of the HSP70 gene in the adenovirus-transformed 293 cell line. Furthermore, the E1A gene was also found to be cell cycle regulated; the activation and peak level of the E1A mRNA occurred at an earlier time than those of the heat shock mRNA, consistent with, but not proof of, the hypothesis that E1A is responsible for the cell cycle control of the HSP70 expression. We therefore suggest that the E1A-like cellular activity may govern certain aspects of cell cycle transcription.

Selective induction of human heat shock gene transcription by the adenovirus E1A gene products, including the 12S E1A product

1987 ◽  
Vol 7 (8) ◽  
pp. 2884-2890
Author(s):  
M C Simon ◽  
K Kitchener ◽  
H T Kao ◽  
E Hickey ◽  
L Weber ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Heat Shock ◽  
Heat Shock Gene ◽  
Hsp70 Expression ◽  
Gene Encoding ◽  
S1 Nuclease ◽  
Adenovirus E1a ◽  
Genes Encoding ◽  
E1a Gene ◽  
The Individual

We have previously shown that the human 70-kilodalton heat shock protein gene (hsp70) is induced by the adenovirus E1A gene product and during the S-G2 phase of the cell cycle. In this study, we investigated the effect of E1A on the expression of other human hsp genes. A gene encoding one form of the hsp89 protein (hsp89 alpha) was activated during an adenovirus infection with kinetics similar to those of activation of hsp70. The induction required a functional E1A gene. However, the hsp89 transcript was not cell cycle regulated. Genes encoding another form of hsp89 and the hsp27 protein were not induced by E1A or during the cell cycle. Further examination of hsp70 expression revealed a greater complexity than previously seen. S1 nuclease analysis using an hsp70 cDNA as well as a distinct hsp70 genomic clone demonstrated three related hsp70 transcripts; two were induced by E1A, and one was not. Both of the E1A-inducible genes were regulated during the cell cycle. All three were induced by heat shock. These results suggest common aspects of control among certain members of this family of cellular genes distinct from heat shock control. Finally, using viruses that express the individual E1A proteins, we found that the hsp70 gene is induced by the 12S and the 13S E1A products. The efficiency of induction by the 12S product was somewhat less than that by the 13S product but only by a factor of less than 2. This is in contrast to the induction of early viral genes, for which the 13S product is considerably more efficient than the 12S product.

scholarly journals Selective induction of human heat shock gene transcription by the adenovirus E1A gene products, including the 12S E1A product.

1987 ◽  
Vol 7 (8) ◽  
pp. 2884-2890 ◽  
Author(s):  
M C Simon ◽  
K Kitchener ◽  
H T Kao ◽  
E Hickey ◽  
L Weber ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Heat Shock ◽  
Heat Shock Gene ◽  
Hsp70 Expression ◽  
Gene Encoding ◽  
S1 Nuclease ◽  
Adenovirus E1a ◽  
Genes Encoding ◽  
E1a Gene ◽  
The Individual

We have previously shown that the human 70-kilodalton heat shock protein gene (hsp70) is induced by the adenovirus E1A gene product and during the S-G2 phase of the cell cycle. In this study, we investigated the effect of E1A on the expression of other human hsp genes. A gene encoding one form of the hsp89 protein (hsp89 alpha) was activated during an adenovirus infection with kinetics similar to those of activation of hsp70. The induction required a functional E1A gene. However, the hsp89 transcript was not cell cycle regulated. Genes encoding another form of hsp89 and the hsp27 protein were not induced by E1A or during the cell cycle. Further examination of hsp70 expression revealed a greater complexity than previously seen. S1 nuclease analysis using an hsp70 cDNA as well as a distinct hsp70 genomic clone demonstrated three related hsp70 transcripts; two were induced by E1A, and one was not. Both of the E1A-inducible genes were regulated during the cell cycle. All three were induced by heat shock. These results suggest common aspects of control among certain members of this family of cellular genes distinct from heat shock control. Finally, using viruses that express the individual E1A proteins, we found that the hsp70 gene is induced by the 12S and the 13S E1A products. The efficiency of induction by the 12S product was somewhat less than that by the 13S product but only by a factor of less than 2. This is in contrast to the induction of early viral genes, for which the 13S product is considerably more efficient than the 12S product.

scholarly journals Posttranscriptional regulation of hsp70 expression in human cells: effects of heat shock, inhibition of protein synthesis, and adenovirus infection on translation and mRNA stability.

1987 ◽  
Vol 7 (12) ◽  
pp. 4357-4368 ◽  
Author(s):  
N G Theodorakis ◽  
R I Morimoto
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Posttranscriptional Regulation ◽  
High Efficiency ◽  
Hsp70 Expression ◽  
Mrna Levels ◽  
Protein Synthesis Inhibitors ◽  
Hsp70 Mrna ◽  
Infected Cells ◽  
The Stability

We have examined the posttranscriptional regulation of hsp70 gene expression in two human cell lines, HeLa and 293 cells, which constitutively express high levels of HSP70. HSP70 mRNA translates with high efficiency in both control and heat-shocked cells. Therefore, heat shock is not required for the efficient translation of HSP70 mRNA. Rather, the main effect of heat shock on translation is to suppress the translatability of non-heat shock mRNAs. Heat shock, however, has a marked effect on the stability of HSP70 mRNA; in non-heat-shocked cells the half-life of HSP70 mRNA is approximately 50 min, and its stability increases at least 10-fold upon heat shock. Moreover, HSP70 mRNA is more stable in cells treated with protein synthesis inhibitors, suggesting that a heat shock-sensitive labile protein regulates its turnover. An additional effect on posttranscriptional regulation of hsp70 expression can be found in adenovirus-infected cells, in which HSP70 mRNA levels decline precipititously late during infection although hsp70 transcription continues unabated.

Transcriptional activation and subsequent control of the human heat shock gene during adenovirus infection

1983 ◽  
Vol 3 (11) ◽  
pp. 2058-2065 ◽  
Author(s):  
H T Kao ◽  
J R Nevins
Keyword(s):  
Heat Shock ◽  
Transcriptional Activation ◽  
Peak Level ◽  
Heat Shock Gene ◽  
Adenovirus Infection ◽  
Wild Type ◽  
Rapid Turnover ◽  
Transformed Cell Line ◽  
Shock Gene ◽  
E1a Gene

A cDNA copy of the major human heat shock mRNA was cloned. The clone is complementary to the mRNA encoding the major 70-kilodalton heat shock protein as shown by hybrid arrest translation. We utilized the cloned DNA to measure induction of the gene during adenovirus infection. The mRNA increases in abundance approximately 100-fold during a wild-type adenovirus infection but does not increase more than 2-fold during an infection in which there is no E1A gene function [high multiplicity of infection of an E1A (-) mutant]. Furthermore, by measuring transcription in isolated nuclei, we found that the induction was transcriptional and was mediated by the E1A gene product. The induction was not maintained, however. After a peak level was obtained, transcription returned to preinfection levels. This decline was also reflected in the cytoplasmic mRNA abundance indicating a rapid turnover of the heat shock mRNA. This rapid turnover of the heat shock mRNA appears to be induced by the viral infection since the heat shock mRNA was found to be stable when synthesized in an adenovirus-transformed cell line.

scholarly journals Heat shock and developmental regulation of the Drosophila melanogaster hsp83 gene.

1989 ◽  
Vol 9 (4) ◽  
pp. 1746-1753 ◽  
Author(s):  
H Xiao ◽  
J T Lis
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Fusion Gene ◽  
Developmental Expression ◽  
Regulatory Sequence ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
High Level ◽  
Hsp83 Gene

In contrast to the hsp70 gene, whose expression is normally at a very low level and increases by more than 2 orders of magnitude during heat shock, the hsp83 gene in Drosophila melanogaster is expressed at high levels during normal development and increases only severalfold in response to heat shock. Developmental expression of the hsp83 gene consists of a high level of tissue-general, basal expression and a very high level of expression in ovaries. We identified regions upstream of the hsp83 gene that were required for its developmental and heat shock-induced expression by assaying beta-galactosidase activity and mRNA levels in transgenic animals containing a series of 5' deletion and insertion mutations of an hsp83-lacZ fusion gene. Deletion of sequences upstream of the overlapping array of a previously defined heat shock consensus (HSC) sequence eliminated both forms of developmental expression of the hsp83 gene. As a result, the hsp83 gene with this deletion mutation was regulated like that of the hsp70 gene. Moreover, an in vivo polymer competition assay revealed that the overlapping HSC sequences of the hsp83 gene and the nonoverlapping HSC sequences of the hsp70 gene had similar affinities for the factor required for heat induction of the two heat shock genes. We discuss the functional similarity of hsp70 and hsp83 heat shock regulation in terms of a revised view of the heat shock-regulatory sequence.

scholarly journals Heat shock results in cell cycle delay and synchronisation of mitotic domains in cellularised Drosophila melanogaster embryos

1993 ◽  
Vol 105 (3) ◽  
pp. 711-720 ◽  
Author(s):  
G. Maldonado-Codina ◽  
S. Llamazares ◽  
D.M. Glover
Keyword(s):  
Cell Cycle ◽  
Heat Shock ◽  
Chromatin Condensation ◽  
Cell Cycle Control ◽  
S Phase ◽  
Temperature Shock ◽  
Cell Cycle Delay ◽  
Minute Period ◽  
G2 Phase ◽  
Drosophila Embryos

Cells of Drosophila embryos that are subjected to a 37 degrees C temperature shock whilst undergoing the S-phase of cell cycle 14 arrest with their microtubules in an interphase-like state, and with nuclei showing unusual chromatin condensation. They do not recover from this state within a 30 minute period even though extensive gastrulation movements can occur. Cells of embryos heat shocked in G2-phase are delayed in interphase with high levels of cyclins A and B. Within ten minutes recovery from heat shock, cells enter mitosis throughout the embryo. The degradation of the mitotic cyclins A and B in these synchronised mitotic domains does not follow the normal timing, but is delayed. These findings point to a need for caution when interpreting experiments that use the heat shock promoter to study the expression of cell cycle control genes in Drosophila.

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.

Heat shock and developmental regulation of the Drosophila melanogaster hsp83 gene

1989 ◽  
Vol 9 (4) ◽  
pp. 1746-1753
Author(s):  
H Xiao ◽  
J T Lis
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Fusion Gene ◽  
Developmental Expression ◽  
Regulatory Sequence ◽  
Mrna Levels ◽  
Hsp70 Gene ◽  
High Level ◽  
Hsp83 Gene

In contrast to the hsp70 gene, whose expression is normally at a very low level and increases by more than 2 orders of magnitude during heat shock, the hsp83 gene in Drosophila melanogaster is expressed at high levels during normal development and increases only severalfold in response to heat shock. Developmental expression of the hsp83 gene consists of a high level of tissue-general, basal expression and a very high level of expression in ovaries. We identified regions upstream of the hsp83 gene that were required for its developmental and heat shock-induced expression by assaying beta-galactosidase activity and mRNA levels in transgenic animals containing a series of 5' deletion and insertion mutations of an hsp83-lacZ fusion gene. Deletion of sequences upstream of the overlapping array of a previously defined heat shock consensus (HSC) sequence eliminated both forms of developmental expression of the hsp83 gene. As a result, the hsp83 gene with this deletion mutation was regulated like that of the hsp70 gene. Moreover, an in vivo polymer competition assay revealed that the overlapping HSC sequences of the hsp83 gene and the nonoverlapping HSC sequences of the hsp70 gene had similar affinities for the factor required for heat induction of the two heat shock genes. We discuss the functional similarity of hsp70 and hsp83 heat shock regulation in terms of a revised view of the heat shock-regulatory sequence.

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