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Expression of leucine zipper-transcription factors in cardiac hypertrophy
88
Cell Biology International
P37
(I): Institut Electron&we, INRA, &ion Josas.
Pasteur, Station 25 rue du Dr de Physiologie
Cei&ale de Microsco$e Roux, 75015 Paris. (21: Animale, 78380 Jouy‘ kn
The vimentin gene is expressed in all cellular types grown in culture and the nucleotide sequence of its regulatory region is highly conserved in different species (human, chicken, mouse). Beside, the T antigen of the simian virus 40 (SV40) is known to confer immortalizing properties on a large variety of cells. We have demonstrated that T antigen stimulates the human vimentin promoter and the activated region contains 9 times the consensus sequence 5!(G/TA/GGGC)3’ recognized by SV40 T antigen. In order to obtain cell lines from primary tissue culture or transgenics animals, we have constructed a recombinant plasmid ( HUPROVIM -T,t ) which contains the early genes of SV40 (T and t antigens) placed under the control of a deletion mutant of the regulatory region of the human vimentin gene. After microinjection of HUPROVIM into the nucleus of primary cells, we have T,t established in culture different cell types: Endothelial cells from human, oorcine and rabbit; Epithelial cells from bovine. - Microinjection of HUPROVIM -T.t into fertilized enas of rabbits were have been achieved and two transgenic &imals derived.
P39
PERIODIC ASSOCIATION OF MYC-PROTEINS WITH THE NUCLEAR MATRIX DURING THE CELL CYCLE Werner Waitz, Peter Loidl. Department of Pathology and Institute for Medical Chemistry & Biochemistry, University of Innsbruck, 6020 Innsbruck, Austria. We demonstrate the presence of c-myc orotein in the lower eukaryot Physarum polycephaium: The protein with Mr 67 kD remains firmlv associated with the nuclear matrix after a variety of matrix isolation procedures. The level of c-myc protein does not significantly fluctuate during the naturally synchronous Physarum cell cycle. However, the matrix-bound portion of c-myc protein is higher during S as commicroscopy the immupared to Gz. In imnunoelectron nosignal is considerably stronger in the nuclear matrix as compared to unfractionated nuclei, indicating increased accessibility of c-myc protein in nuclear scaffold structures. We demonstrate that the nuclear localization of c-myc protein changes during the cell cycle. It is transiently, but specifically bound to the nuclear lamina during S-phase, probably reflecting an intimate role of this protein in DNAreplication. In Gp-period the immunosignal is distributed evenly over the whole nuclear matrix. The results indicate that the participation of mycprotein in cell-cycle regulated processes might be modulated by a periodic association to a nuclear substructure, rather than by the overall amount of the protein.
1990
EXPRESSIDR OF LEUCINE ZIPPER-TRARXRIPTION FACTORS IN CARDIAC RYPERTROPNY Thomas Brand, Hari S Sharma. Piero A Martorana*. Wolfgang Schaper. MPI Bad Nauheim, Exp. Cardiology, 6350 Bad Nauheim. FRG. Cassella Aa. De&. of Pharmacology, 6000‘Frankfurt, FRG*. "' ' Cardiac hypertrophy is a multistep process which leads to adaptation of the myocardium to increased wall stress. Hypertrophied myocytes have an altered pattern of gene expression: e.g. isoform switches of contractile proteins, reeinduction of embrvonic genes. Proto-oncogenes.like c-fos, c-myc and c-jun are DNA-bindinq oroteins which reaulate transcription. Thejr form dimers with the"help of the so-called leucine zipper structure. We studied the expression of several leucine zipper genes in cardiac hypertrophy. Hypertrophy was induced in adult rats by infusion of the beta-adrenergic agonist isoproterenol (iso). In the first 24 h an induction of c-myc, c-fos and fos-B mRNA was observed. However basal expression of c-jun was not enhanced. We also observed an induction of hsp-70, the major heat shock protein. Blockade of the beta receptor by propanolol prevented the induction of c-fos. Injection of dBcAMP induced c-fos, but to a much lower level compared to iso infusion. Right ventricular hypertrophy is a known consequence of lung emphysema. The tsk mouse a genetic model of lung emphysema develops right ventricular hvpertroohv which becomes aooarent at the aae of 16 month. We detected a strong signal for E-fos in riqht ventricle but not in left ventricle of 16 month old tsk mouse, wheras in 8 month old tsk mouse no expression of c-fos was observed. These results imply a relationship between induction of leucine zipper - transcription factors and the altered gene expression in hypertrophied heart.
P38
IN VITRO IMMORTALIZATION: SUPPRESSION OF SPECIES AND CELL-TYPE RESTRICTIONS BY USE OF SIMIAN VIRUS 40 T ANTIGEN UNDER THECONTROLOFARRGULATORYREGIONOFTHE VIMENTIN GENE. Patrick VICART(l), Bertrand SCHWARTZQ). Claude DELOUIS(2). Denise PAULIN(
Reports, Vol. 14, Abstracts Supplement
Cell Biology International
P37
(I): Institut Electron&we, INRA, &ion Josas.
Pasteur, Station 25 rue du Dr de Physiologie
Cei&ale de Microsco$e Roux, 75015 Paris. (21: Animale, 78380 Jouy‘ kn
The vimentin gene is expressed in all cellular types grown in culture and the nucleotide sequence of its regulatory region is highly conserved in different species (human, chicken, mouse). Beside, the T antigen of the simian virus 40 (SV40) is known to confer immortalizing properties on a large variety of cells. We have demonstrated that T antigen stimulates the human vimentin promoter and the activated region contains 9 times the consensus sequence 5!(G/TA/GGGC)3’ recognized by SV40 T antigen. In order to obtain cell lines from primary tissue culture or transgenics animals, we have constructed a recombinant plasmid ( HUPROVIM -T,t ) which contains the early genes of SV40 (T and t antigens) placed under the control of a deletion mutant of the regulatory region of the human vimentin gene. After microinjection of HUPROVIM into the nucleus of primary cells, we have T,t established in culture different cell types: Endothelial cells from human, oorcine and rabbit; Epithelial cells from bovine. - Microinjection of HUPROVIM -T.t into fertilized enas of rabbits were have been achieved and two transgenic &imals derived.
P39
PERIODIC ASSOCIATION OF MYC-PROTEINS WITH THE NUCLEAR MATRIX DURING THE CELL CYCLE Werner Waitz, Peter Loidl. Department of Pathology and Institute for Medical Chemistry & Biochemistry, University of Innsbruck, 6020 Innsbruck, Austria. We demonstrate the presence of c-myc orotein in the lower eukaryot Physarum polycephaium: The protein with Mr 67 kD remains firmlv associated with the nuclear matrix after a variety of matrix isolation procedures. The level of c-myc protein does not significantly fluctuate during the naturally synchronous Physarum cell cycle. However, the matrix-bound portion of c-myc protein is higher during S as commicroscopy the immupared to Gz. In imnunoelectron nosignal is considerably stronger in the nuclear matrix as compared to unfractionated nuclei, indicating increased accessibility of c-myc protein in nuclear scaffold structures. We demonstrate that the nuclear localization of c-myc protein changes during the cell cycle. It is transiently, but specifically bound to the nuclear lamina during S-phase, probably reflecting an intimate role of this protein in DNAreplication. In Gp-period the immunosignal is distributed evenly over the whole nuclear matrix. The results indicate that the participation of mycprotein in cell-cycle regulated processes might be modulated by a periodic association to a nuclear substructure, rather than by the overall amount of the protein.
1990
EXPRESSIDR OF LEUCINE ZIPPER-TRARXRIPTION FACTORS IN CARDIAC RYPERTROPNY Thomas Brand, Hari S Sharma. Piero A Martorana*. Wolfgang Schaper. MPI Bad Nauheim, Exp. Cardiology, 6350 Bad Nauheim. FRG. Cassella Aa. De&. of Pharmacology, 6000‘Frankfurt, FRG*. "' ' Cardiac hypertrophy is a multistep process which leads to adaptation of the myocardium to increased wall stress. Hypertrophied myocytes have an altered pattern of gene expression: e.g. isoform switches of contractile proteins, reeinduction of embrvonic genes. Proto-oncogenes.like c-fos, c-myc and c-jun are DNA-bindinq oroteins which reaulate transcription. Thejr form dimers with the"help of the so-called leucine zipper structure. We studied the expression of several leucine zipper genes in cardiac hypertrophy. Hypertrophy was induced in adult rats by infusion of the beta-adrenergic agonist isoproterenol (iso). In the first 24 h an induction of c-myc, c-fos and fos-B mRNA was observed. However basal expression of c-jun was not enhanced. We also observed an induction of hsp-70, the major heat shock protein. Blockade of the beta receptor by propanolol prevented the induction of c-fos. Injection of dBcAMP induced c-fos, but to a much lower level compared to iso infusion. Right ventricular hypertrophy is a known consequence of lung emphysema. The tsk mouse a genetic model of lung emphysema develops right ventricular hvpertroohv which becomes aooarent at the aae of 16 month. We detected a strong signal for E-fos in riqht ventricle but not in left ventricle of 16 month old tsk mouse, wheras in 8 month old tsk mouse no expression of c-fos was observed. These results imply a relationship between induction of leucine zipper - transcription factors and the altered gene expression in hypertrophied heart.
P38
IN VITRO IMMORTALIZATION: SUPPRESSION OF SPECIES AND CELL-TYPE RESTRICTIONS BY USE OF SIMIAN VIRUS 40 T ANTIGEN UNDER THECONTROLOFARRGULATORYREGIONOFTHE VIMENTIN GENE. Patrick VICART(l), Bertrand SCHWARTZQ). Claude DELOUIS(2). Denise PAULIN(
Reports, Vol. 14, Abstracts Supplement