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

• Autorzy: Widlak W. , Vydra N. , Dudaladava V. , Scieglinska D. , Winiarski B. , Krawczyk Z.
• Języki publikacji: EN

Abstrakty

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.

Słowa kluczowe

EN

gene; Hsp70.2-Hst70 gene; gene expression; regulation; spermatozoon; heat shock protein; spermatogenesis; testis-specific gene

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2007
• Tom: 54
• Numer: 1
• Opis fizyczny: p.107-112,fig.,ref.

Twórcy

autor

Widlak W.

• Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Gliwice, Poland

autor

Vydra N.

autor

Dudaladava V.

autor

Scieglinska D.

autor

Winiarski B.

autor

Krawczyk Z.

Bibliografia

• Allen RL, O’Brien DA, Jones CC, Rockett DL, Eddy EM (1988) Expression of heat shock proteins by isolated mouse spermatogenic cells. Mol Cell Biol 8: 3260–3266.
• Bonny Ch, Cooker LA, Goldberg E (1998) Deoxyribonucleic acid-protein interactions and expression of the human testis-specific lactate dehydrogenase promoter: transcription factor Spl plays a major role. Biol Reprod 58: 754–759.
• Bonnycastle LLC, Yu Ch-E, Hunt CR, Trask BJ, Clancy KP, Weber JL, Patterson D, Schellenberg GD (1994) Cloning, sequencing, and mapping of the human chromosome 14 heat shock protein gene (HspA2 ). Genomics 23: 85–93.
• Dix DJ, Allen JW, Collins W, Mori C, Nakamura N, Poorman- Allen P, Goulding EH, Eddy EM (1996) Targeted gene disruption of Hsp70-2 results in failed meiosis, germ cell apoptosis, and male infertility. Proc Natl Acad Sci USA 16: 3264–3268.
• Gebara MM, McCarrey JR (1992) Protein–DNA interactions associated with the onset of testis-specific expression of the mammalian Pgk-2 gene. Mol Cell Biol 12: 1422– 1431.
• Grimes SR, Wolfe SA, Koppel DA (1992) Tissue-specific binding of testis nuclear proteins to a sequence element within the promoter of the testis-specific histone H1t gene. Arch Biochem Biophys 296: 402–409.
• Iannello RC, Kola I, Dahl HH (1993) Temporal and tissuespecific interactions involving novel transcription factors and the proximal promoter of the mouse Phda-2 gene. J Biol Chem 268: 22581–22590.
• Iannello RC, Young J, Sumarsono S, Tymms MJ, Dahl HH, Gould J, Hedger M, Kola I (1997) Regulation of Phda-2 expression is mediated by proximal promoter sequences and CpG methylation. Mol Cell Biol 17: 612–619.
• Kaczynski J, Cook T, Urrutia R (2003) Sp1- and Krüppellike transcription factors. Genome Biol 4: 206.
• Kadonaga JT, Carner KR, Masiarz FR, Tjian R (1987) Isolation of cDNA encoding transcription factor SP1 and functional analysis of the DNA binding domain. Cell 51: 1079–1090.
• Krawczyk Z, Wiśniewski J, Biesiada E (1988a) A rat testis- specific hsp70 gene-related transcript is coded by a novel gene from the hsp70 multigene family. Acta Biochim Polon 35: 377–385.
• Krawczyk Z, Mali P, Parvinen M (1988b) Expression of testis- specific hsp70 gene-related RNA in defined stages of rat seminiferous epithelium. J Cell Biol 107: 1317–1323.
• Krawczyk Z, Schmid W, Harkonen P, Wolniczek P (1993) The ERE-like sequence from the promoter region of the testis specific hsp70-related gene is not estrogen responsive. Cell Biol Int 17: 245–253.
• Persengiev SP, Raval PJ, Rabinovitch S, Millette CF, Kilpatrick DL (1996) Transcription factor Sp1 is expressed by three different developmentally regulated messenger ribonucleic acids in mouse spermatogenic cells. Endocrinology 137: 638–646.
• Pothier F, Ouellet M, Julien JP, Guerin SL (1992) An improved CAT assay for promoter analysis in either transgenic mice or tissue culture cells. DNA Cell Biol 11: 83–90.
• Robidoux S, Gosselin P, Harvey M, Leclerc S, Guerin SL (1992) Transcription of the mouse secretory protease inhibitor p12 gene is activated by the developmentally regulated positive transcription factor Spl. Mol Cell Biol 12: 3796–3806.
• Saffer JD, Jackson SP, Annarella MB (1991) Developmental expression of Spl in the mouse. Mol Cell Biol 11: 2189– 2199.
• Ścieglinska D, Widłak W, Konopka W, Poutanen M, Rahman N, Huhtaniemi I, Krawczyk Z (2001) Structure of the 5’ region of the hst70 gene transcription unit presence of an intron and multiple transcription initiation sites. Biochem J 359: 129–137.
• Ścieglińska D, Vydra N, Krawczyk Z, Widłak W (2004) Location of promoter elements necessary and sufficient to direct testis-specific expression of the Hst70/Hsp70.2 gene. Biochem J 379: 739–747.
• Thomas K, Sung D-Y, Yang J, Johnson K, Thompson W, Millette C, McCarrey J, Breitberg A, Gibbs R, Walker W (2005) Identification, characterization and functional analysis of Sp1 transcript variants expressed in germ cells during mouse spermatogenesis. Biol Reprod 72: 898–907.
• Widłak W, Markkula M, Krawczyk Z, Huhtaniemi I (1994) Functional analysis of spermatocyte-specific hst70 gene promoter in transgenic mice. Acta Biochim Polon 41: 103–105.
• Widłak W, Markkula M, Krawczyk Z, Kananen K, Huhtaniemi I (1995) A 252 bp upstream region of the rat spermatocyte-specific hst70 gene is sufficient to promote expression of the hst70-CAT hybrid gene in testis and brain of transgenic mice. Biochim Biophys Acta 1264: 191–200.
• Widłak W, Ścieglińska D, Vydra N, Małusecka E, Krawczyk Z (2003) In vivo electroporation of the testis versus transgenic mice model in functional studies of spermatocyte- specific hst70 gene promoter. A comparative study. Mol Reprod Dev 65: 382–388.
• Wilkerson DC, Wolfe SA, Grimes SR (2002) H1t/GC-box and H1t/TE1 elements are essential for promotor activity of the testis-specific histone H1t gene. Biol Reprod 67: 1157–1164.
• Zakeri ZF, Wolgemuth DJ, Hunt CR (1988) Identification and sequence analysis of a new member of the mouse HSP70 gene family and characterization of its unique cellular and developmental pattern of expression in the male germ line. Mol Cell Biol 8: 2925–2932.