Application of heat shock promoter in transgenic zebrafish

Heat shock inductors have potential as treatment for degenerative and protein misfolding diseases. Dimethyl-sulfoxide is widely used as a solvent in pharmacological screening tests and has been shown to have heat shock induction effects. Transgenic Tg (hsp70l:EGFP-HRAS_G12V)io3(AB) zebrafish larvae were exposed for 24 hours to dimethyl-sulfoxide in concentratios of 0.1-2%, and to moderate heat shock inductors pentoxifylline and tacrolimus. Positive controls were exposed to 35, 38 and 40�C for 20 min, and incubated for 24 h at 28�C. Heat shock response was measured by fluorescence microscopy and signal intensity quantification in FIJI. Dimethyl-sulfoxide caused a dose-dependant increase in fluorescent intensity, but significantly lower compared with exposure to 38 and 40�C. Pentoxifylline and tacrolimus induced a significantly higher increase in fluorescence compared with 0.5% dimethyl-sulfoxide. Thus, although dimethyl-sulfoxide has independent heat shock induction effects, concentrations of up to 0.5% are suitable for heat shock response screening tests.

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A predicted membrane protein, TRA-2A, directs hermaphrodite development in Caenorhabditis elegans

Development ◽

10.1242/dev.121.9.2995 ◽

1995 ◽

Vol 121 (9) ◽

pp. 2995-3004 ◽

Cited By ~ 7

Author(s):

P.E. Kuwabara ◽

J. Kimble

Keyword(s):

Heat Shock ◽

Membrane Protein ◽

Cell Fate ◽

Loss Of Function ◽

Heat Shock Promoter ◽

Somatic Development ◽

C Elegans ◽

Cellular Domain ◽

Carboxy Terminal ◽

Necessary And Sufficient

The nematode C. elegans naturally develops as either an XO male or XX hermaphrodite. The sex-determining gene, tra-2, promotes hermaphrodite development in XX animals. This gene encodes a predicted membrane protein, named TRA-2A, which has been proposed to provide the primary feminising activity of the tra-2 locus. Here, we show that transgenic TRA-2A driven from a heat shock promoter can fully feminise the somatic tissues of XX tra-2 loss-of-function mutants, which would otherwise develop as male. TRA-2A is thus likely to provide a component of the tra-2 locus that is both necessary and sufficient to promote female somatic development. Transgenic TRA-2A driven by the heat shock promoter can also transform XO animals from male to self-fertile hermaphrodite. This result establishes the role of tra-2 as a developmental switch that controls somatic sexual cell fate. We show that a carboxy-terminal region of TRA-2A, predicted to be intra-cellular, can partially feminise XX tra-2 loss-of-function mutants and XO tra-2(+) males. We suggest that this intra-cellular domain of TRA-2A promotes hermaphrodite development by negatively regulating the FEM proteins.

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Ultrasound-Induced Expression of a Heat Shock Promoter-Driven Transgene Delivered in the Kidney by Genetically Modified Mesenchymal Stem Cells

MRI-Guided Focused Ultrasound Surgery ◽

10.3109/9781420019933-15 ◽

2007 ◽

pp. 171-179 ◽

Cited By ~ 1

Author(s):

Béatrice Letavernier ◽

Rares Salomir ◽

Yahsou Delmas ◽

Claire Rome ◽

Franck Couillaud ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Heat Shock ◽

Genetically Modified ◽

Induced Expression ◽

Heat Shock Promoter

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Identification of a heat shock promoter in the topA gene of Escherichia coli.

Journal of Bacteriology ◽

10.1128/jb.172.12.6871-6874.1990 ◽

1990 ◽

Vol 172 (12) ◽

pp. 6871-6874 ◽

Cited By ~ 22

Author(s):

S A Lesley ◽

S B Jovanovich ◽

Y C Tse-Dinh ◽

R R Burgess

Keyword(s):

Escherichia Coli ◽

Heat Shock ◽

Heat Shock Promoter

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P transposons controlled by the heat shock promoter.

Molecular and Cellular Biology ◽

10.1128/mcb.6.5.1640 ◽

1986 ◽

Vol 6 (5) ◽

pp. 1640-1649 ◽

Cited By ~ 50

Author(s):

H Steller ◽

V Pirrotta

Keyword(s):

Heat Shock ◽

Germ Line ◽

Fusion Gene ◽

P Element ◽

Translational Efficiency ◽

Heat Shock Promoter ◽

P Elements ◽

Heat Shocks ◽

Neomycin Resistance ◽

Heat Shock Induction

We have transformed Drosophila melanogaster with modified P-element transposons, which express the transposase function from the heat-inducible hsp70 heat shock promoter. The Icarus transposon, which contains a direct hsp70-P fusion gene, behaved like a very active autonomous P element even before heat shock induction. Although heat shock led to abundant somatic transcription, transposition of the Icarus element was confined to germ line cells. To reduce the constitutive transposase activity observed for the Icarus element, we attenuated the translational efficiency of the transposase RNA by inserting the transposon 5 neomycin resistance gene between the hsp70 promoter and the P-element sequences. The resulting construct, called Icarus-neo, conferred resistance to G418, and its transposition was significantly stimulated by heat shock. Heat shocks applied during the embryonic or third instar larval stage had similar effects, indicating that transposition of P elements is not restricted to a certain developmental stage. Both Icarus and Icarus-neo destabilized snw in a P-cytotype background and thus at least partially overcome the repression of transposition. Our results suggest that the regulation of P-element transposition occurs at both the transcriptional and posttranscriptional levels.

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Induction of dnaK through Its Native Heat Shock Promoter Is Necessary for Intramacrophagic Replication of Brucella suis

Infection and Immunity ◽

10.1128/iai.70.3.1631-1634.2002 ◽

2002 ◽

Vol 70 (3) ◽

pp. 1631-1634 ◽

Cited By ~ 27

Author(s):

Stephan Köhler ◽

Euloge Ekaza ◽

Jean-Yves Paquet ◽

Karl Walravens ◽

Jacques Teyssier ◽

...

Keyword(s):

Heat Shock ◽

Mouse Model ◽

High Sensitivity ◽

Murine Macrophage ◽

Null Mutant ◽

Intracellular Replication ◽

Heat Shock Promoter ◽

Brucella Suis ◽

Independent Synthesis ◽

Mouse Model Of Infection

ABSTRACT The heat shock protein DnaK is essential for intramacrophagic replication of Brucella suis. The replacement of the stress-inducible, native dnaK promoter of B. suis by the promoter of the constitutively expressed bla gene resulted in temperature-independent synthesis of DnaK. In contrast to a dnaK null mutant, this strain grew at 37°C, with a thermal cutoff at 39°C. However, the constitutive dnaK mutant, which showed high sensitivity to H2O2-mediated stress, failed to multiply in murine macrophage-like cells and was rapidly eliminated in a mouse model of infection, adding strong arguments to our hypothesis that stress-mediated and heat shock promoter-dependent induction of dnaK is a crucial event in the intracellular replication of B. suis.

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The DNA-binding activity of the human heat shock transcription factor is regulated in vivo by hsp70.

Molecular and Cellular Biology ◽

10.1128/mcb.13.9.5427 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5427-5438 ◽

Cited By ~ 144

Author(s):

D D Mosser ◽

J Duchaine ◽

B Massie

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Dna Binding ◽

Elevated Temperatures ◽

Active Role ◽

Heat Shock Transcription Factor ◽

Response To Treatment ◽

Promoter Element ◽

Heat Shock Promoter

The human heat shock transcription factor (HSF) is maintained in an inactive non-DNA-binding form under nonstress conditions and acquires the ability to bind specifically to the heat shock promoter element in response to elevated temperatures or other conditions that disrupt protein structure. Here we show that constitutive overexpression of the major inducible heat shock protein, hsp70, in transfected human cells reduces the extent of HSF activation after a heat stress. HSF activation was inhibited more strongly in clones that express higher levels of hsp70. These results demonstrate that HSF activity is negatively regulated in vivo by hsp70 and suggest that the cell might sense elevated temperature as a decreased availability of hsp70. HSF activation in response to treatment with sodium arsenite or the proline analog azetidine was also depressed in hsp70-expressing cells relative to that in the nontransfected control cells. As well, the level of activated HSF decreased more rapidly in the hsp70-expressing clones when the cells were heat shocked and returned to 37 degrees C. These results suggest that hsp70 could play an active role in the conversion of HSF back to a conformation that does not bind the heat shock promoter element during the attenuation of the heat shock response.

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