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1

Functional analysis of spermatocyte-specific hst70 gene promoter in transgenic mice

100%

Widłak W., Markkula M., Krawczyk Z., Huhtaniemi I.

Acta Biochimica Polonica

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1994

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vol. 41

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issue 2

103-105

2

Isolation and structural analysis of rat heat-inducible hsp70 gene

88%

Lisowska K., Krawczyk Z., Widłak W., Wolniczek P.

Acta Biochimica Polonica

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1994

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vol. 41

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issue 2

110-112

3

The GC-box is critical for high level expression of the testis-specific Hsp70.2/Hst70 gene

76%

Widłak W., Vydra N., Dudaladava V., Ścieglińska D., Winiarski B., Krawczyk Z.

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2007

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vol. 54

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issue 1

107-112

EN

The Hsp70.2/Hst70 gene, which belongs to the 70 kDa heat-shock protein (HSP) family, is expressed specifically in primary spermatocytes and spermatids. The regulatory elements required for a high level of testis-specific expression of the gene are placed between the two major transcription start sites T1 and T2 (approximately 350 and 115 bp upstream of the starting ATG codon). Here we have shown that sequences proximal to the exon1/intron splicing site in the 5' untranslated region of the Hsp70.2/Hst70 gene, which include a highly conserved element called box B, are required for efficient expression of the chloramphenicol acetyltransferase reporter gene in testes of transgenic mice. However, in spite of the drastically reduced overall activity, the stage-specific expression pattern of the transgene was preserved after removal of these sequences. We have also shown that GC-box located downstream of the box B (approximately 210 bp upstream of the starting ATG codon) is indispensable for efficient expression of the Hsp70.2/Hst70 gene promoter in spermatogenic cells. The GC-box specifically binds proteins present in nuclear extracts from testes (putatively Sp1-like factors). A change in the pattern of such GC-box-interacting factors corresponds to activation of the Hsp70.2/Hst70 gene, confirming the importance of this regulatory element.

4

Expression of a constitutively active mutant of heat shock factor 1 under the control of testis-specific hst70 gene promoter in transgenic mice induces degeneration of seminiferous epithelium.

64%

Widłak W., Benedyk K., Vydra N., Głowala M., Ścieglińska D., Małusecka E., Nakai A., Krawczyk Z.

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2003

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vol. 50

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issue 2

535-541

EN

Heat shock activates in somatic cells a set of genes encoding heat shock proteins which function as molecular chaperones. The basic mechanism by which these genes are activated is the interaction of the specific transcription factor HSF1 with a regulatory DNA sequence called heat shock element (HSE). In higher eukaryotes HSF1 is present in unstressed cells as inactive monomers which, in response to cellular stress, aggregate into transcriptionally competent homotrimers. In the present paper we showed that the expression of a transgene encoding mutated constitutively active HSF1 placed under the control of a spermatocyte-specific promoter derived from the hst70 gene severely affects spermatogenesis. We found the testes of transgenic mice to be significantly smaller than those of wild-type males and histological analysis showed massive degeneration of the seminiferous epithelium. The lumen of tubules was devoid of spermatids and spermatozoa and using the TUNEL method we demonstrated a high rate of spermatocyte apoptosis. The molecular mechanism by which constitutively active HSF1 arrests spermatogenesis is not known so far. One can assume that HSF1 can either induce or repress so far unknown target genes involved in germ cell apoptosis.

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4 Acta Biochimica Polonica

1 2007

1 2003

2 1994

4 Krawczyk Z.

4 Widłak W.

2 Vydra N.

2 Ścieglińska D.

1 Benedyk K.

1 Dudaladava V.

1 Głowala M.

1 Huhtaniemi I.

1 Lisowska K.

1 Markkula M.

1 Małusecka E.

1 Nakai A.

1 Winiarski B.

1 Wolniczek P.

Functional analysis of spermatocyte-specific hst70 gene promoter in transgenic mice

Isolation and structural analysis of rat heat-inducible hsp70 gene

The GC-box is critical for high level expression of the testis-specific Hsp70.2/Hst70 gene

Expression of a constitutively active mutant of heat shock factor 1 under the control of testis-specific hst70 gene promoter in transgenic mice induces degeneration of seminiferous epithelium.