Jan Mous - Academia.edu (original) (raw)
Papers by Jan Mous
Gene Therapy, 1997
Amplicon vectors incorporate genetic elements from Herpes simplex virus (HSV) in a plasmid form w... more Amplicon vectors incorporate genetic elements from Herpes simplex virus (HSV) in a plasmid form which is packaged into virions in the presence of a replication-defective helper virus. We constructed a new amplicon vector, pHermes-tet-lacZ, that carries the bacterial beta-galactosidase (lacZ) gene under the control of a minimal promoter preceded by a heptameric tetracycline operator. The minimal promoter element is activated by a tetracycline-responsive hybrid protein, the gene for which is also present in the vector. This amplicon was propagated in parallel in two different permissive cell lines, E5 and 2-2, in the presence of two helper viruses, d120 and 5dl1.2, respectively. The viral stocks produced were injected into the hippocampal region of the mouse brain, where strong localized expression of the transgene developed in the granular cell layer of the dentate gyrus with limited cytotoxicity. The transgene expression could be repressed by a factor of 10 after administration of tetracyclines. The repression level depended on the helper virus present in the injected viral stock. The in vivo regulation of transgene expression conferred by the tetracycline responsive element improves the flexibility of amplicon vectors as tools for gene transfer into the brain.
Molecular and Cellular Biology, 2003
Nuclear receptors are ligand-modulated transcription factors. On the basis of the completed human... more Nuclear receptors are ligand-modulated transcription factors. On the basis of the completed human genome sequence, this family was thought to contain 48 functional members. However, by mining human and mouse genomic sequences, we identified FXR as a novel family member. It is a functional receptor in mice, rats, rabbits, and dogs but constitutes a pseudogene in humans and primates. Murine FXR is widely coexpressed with FXR in embryonic and adult tissues. It heterodimerizes with RXR␣ and stimulates transcription through specific DNA response elements upon addition of 9-cis-retinoic acid. Finally, we identified lanosterol as a candidate endogenous ligand that induces coactivator recruitment and transcriptional activation by mFXR.
Gene, 1987
95-103 Elsevier GEN 02033 95 Expression of biologically active human T-cell lymphotropic virus ty... more 95-103 Elsevier GEN 02033 95 Expression of biologically active human T-cell lymphotropic virus type III reverse transcriptase in Bacillus subtilis (Repressible expression system; recombinant DNA; immunoblotting; human T-cell; lac operator; phage T5 promoter; protease) SUMMARY A 2.4-kb DNA fragment from the pol region of the human T-cell lymphotropic virus, encoding the protease, reverse transcriptase and a portion of the endonuclease N-terminus was stably introduced into a high-level Bacillus subtilis expression system under inducible control of the Escherichia coli lac regulatory elements. The major expression plasmid, pRTLl1, contains a bacteriophage T5 promoter/lac operator element, which is controlled by lac repressor, supplied by the secondary plasmid, pBL1. Upon IPTG induction, a 90-kDa polyprotein is synthesised and subsequently proteolytically cleaved to reveal 64-kDa and 52-kDa polypeptides. Partial purification reveals that reverse transcriptase activity co-migrates with these two polypeptides.
Nature Genetics, 2000
Many pathological processes, including those causing allergies and autoimmune diseases, are assoc... more Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation 1 . Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes 2 . Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells.
Nature Genetics, 2000
Many pathological processes, including those causing allergies and autoimmune diseases, are assoc... more Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation 1 . Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes 2 . Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells.
Nature Biotechnology, 1998
DNA Arrays, 2001
ABSTRACT Miniaturization and high-throughput parallel analysis are new concepts entering modern m... more ABSTRACT Miniaturization and high-throughput parallel analysis are new concepts entering modern molecular biology. An exciting example of such technology originally developed by physicists and applied to biology is the microchip. The semiconductor industry manufactures silica chips with increasing numbers of smaller and smaller features, which allows incredible numbers of operations within split seconds. The same principle of performing multiparallel operations on a miniaturized solid phase has led to the development of miniaturized arrays of several hundreds of thousands of DNA fragments on a small chip. Arraying of DNA or RNA samples onto nitrocellulose or nylon membranes is a very common analytical procedure in a molecular biology laboratory. DNA chips or microarrays are miniaturized versions of these classical filter-hybridization techniques. With great precision, thousands of DNA fragments or DNA oligonucleotides can be printed onto glass chips or microscope slides. In the case of oligonucleotides, they can also be synthesized directly on coated silica chips.
Nature Biotechnology, 1998
Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1979
1. Total RNA was extracted from human term placenta and mRNA purified by chromatography on oligo(... more 1. Total RNA was extracted from human term placenta and mRNA purified by chromatography on oligo(dT)-cellulose. The poly(A)-containing fraction stimulated amino acid incorporation 5- to 10-fold in the wheat germ cell-free system. Immunoprecipitation with an anti-lactogen serum indicated that 14-27% of the peptides synthesized in vitro contained antigenic determinants of this hormone. 2. Analysis of the [3H]leucine labelled product in the immunoprecipitate on sodium dodecyl sulfate-polyacrylamide gels revealed a complex mixture of polypeptides. Two heavily labelled bands (I and III) were seen corresponding in mobility with pre-lactogen (Mr = 25 000) and native lactogen (Mr = 22 200), each accounting for about 30% of the immunoprecipitable radioactivity. Two additional bands with an intermediate mobility were also observed. 3. Synthesis of the hormone was inhibited by 7-methylguanosine-5'-monophosphate suggesting the presence of a 7-methylguanosine 'cap' on the 5'-end of the mRNA for lactogen. 4. Peptide analysis of the cyanogen bromide cleavage products of band I, band III and authentic lactogen showed marked similarities in their primary structure. The precursor molecule, however, was lacking the N-terminal peptide present in authentic hormone indicating the presence of an extension of 25 amino acids at this side of the molecule. 5. The presence of one or several processing enzymes in the wheat germ cell-free system was indicated by the effect of Triton X-100. Low concentrations of this detergent (0.04%) while inhibiting the protein synthesizing activity for only 15%, completely abolished the precursor cleavage activity. Under these conditions only pre-lactogen was detected in the immunoprecipitate.
European Journal of Biochemistry, 1982
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1983
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1982
The amino acid sequence of component C1, the polypeptide specific for subunit F of prostatic bind... more The amino acid sequence of component C1, the polypeptide specific for subunit F of prostatic binding protein, the major secretory glycoprotein of the rat ventral prostate, has been determined. Its structure was established using the manual Edman degradation on the intact protein and on the most relevant fragments isolated from trypsin, chymotrypsin, thermolysin and Staphylococcus aureus protease digests of the 14C-labelled S-carboxamidornethylated component C1.Component C1 contains 88 amino acids corresponding to a molecular weight of 10246. It is an acidic polypeptide due to the presence of 17 acidic residues; its thrce cysteine residues are almost symmetrically distributed over the peptide chain. Highly polar regions are found in positions 17-27 and 37-47, while the C-terminal part of the molecule contains two hydrophobic segments
European Journal of Biochemistry, 1978
The prostatic binding protein, previously described in rat ventral prostate, was isolated. The pu... more The prostatic binding protein, previously described in rat ventral prostate, was isolated. The purified protein binds pregnenolone with an affinity of 1.2 × 106 M−1 and contains an average of 0.84 binding site per molecule. Its carbohydrate content is 3.2%. Its Mr, estimated by gel filtration, is 51000 but in the presence of 6 M guanidine hydrochloride or 0.1% dodecylsufate it dissociates into two subunits (S and F), which can be separated by polyacrylamide gel electrophoresis or by chromatography on hydroxyapatite. The Mr of these subunits is about 17000, when estimated by gel filtration in 6 M guanidine hydrochloride, or 19000 for subunit F and 20000 for subunit S, when measured by dodecylsulfate/polyacrylamide gel electrophoresis. Their isoelectric points, estimated by isoelectric focusing in 8 M urea, are 4.6 for subunit F and 4.9 for subunit S. Prostatic binding protein and both subunits have a very similar amino acid composition. Upon reduction of disulfide bridges each subunit dissociates further into two components: one of these components is the same in both subunits.
European Journal of Biochemistry - EUR J BIOCHEM, 2005
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1978
European Journal of Biochemistry, 1983
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1979
Polysomal RNA was extracted from human term placenta and total poly(A)-containing RNA purified by... more Polysomal RNA was extracted from human term placenta and total poly(A)-containing RNA purified by affinity chromatography on oligo(dT)-cellulose. Poly(A)-containing RNA constituted approximately 1.2 "/, of the total polysomal RNA and 8 % of this purified preparation was able to anneal with [3H
European Journal of Biochemistry, 1982
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 2005
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1983
Gene Therapy, 1997
Amplicon vectors incorporate genetic elements from Herpes simplex virus (HSV) in a plasmid form w... more Amplicon vectors incorporate genetic elements from Herpes simplex virus (HSV) in a plasmid form which is packaged into virions in the presence of a replication-defective helper virus. We constructed a new amplicon vector, pHermes-tet-lacZ, that carries the bacterial beta-galactosidase (lacZ) gene under the control of a minimal promoter preceded by a heptameric tetracycline operator. The minimal promoter element is activated by a tetracycline-responsive hybrid protein, the gene for which is also present in the vector. This amplicon was propagated in parallel in two different permissive cell lines, E5 and 2-2, in the presence of two helper viruses, d120 and 5dl1.2, respectively. The viral stocks produced were injected into the hippocampal region of the mouse brain, where strong localized expression of the transgene developed in the granular cell layer of the dentate gyrus with limited cytotoxicity. The transgene expression could be repressed by a factor of 10 after administration of tetracyclines. The repression level depended on the helper virus present in the injected viral stock. The in vivo regulation of transgene expression conferred by the tetracycline responsive element improves the flexibility of amplicon vectors as tools for gene transfer into the brain.
Molecular and Cellular Biology, 2003
Nuclear receptors are ligand-modulated transcription factors. On the basis of the completed human... more Nuclear receptors are ligand-modulated transcription factors. On the basis of the completed human genome sequence, this family was thought to contain 48 functional members. However, by mining human and mouse genomic sequences, we identified FXR as a novel family member. It is a functional receptor in mice, rats, rabbits, and dogs but constitutes a pseudogene in humans and primates. Murine FXR is widely coexpressed with FXR in embryonic and adult tissues. It heterodimerizes with RXR␣ and stimulates transcription through specific DNA response elements upon addition of 9-cis-retinoic acid. Finally, we identified lanosterol as a candidate endogenous ligand that induces coactivator recruitment and transcriptional activation by mFXR.
Gene, 1987
95-103 Elsevier GEN 02033 95 Expression of biologically active human T-cell lymphotropic virus ty... more 95-103 Elsevier GEN 02033 95 Expression of biologically active human T-cell lymphotropic virus type III reverse transcriptase in Bacillus subtilis (Repressible expression system; recombinant DNA; immunoblotting; human T-cell; lac operator; phage T5 promoter; protease) SUMMARY A 2.4-kb DNA fragment from the pol region of the human T-cell lymphotropic virus, encoding the protease, reverse transcriptase and a portion of the endonuclease N-terminus was stably introduced into a high-level Bacillus subtilis expression system under inducible control of the Escherichia coli lac regulatory elements. The major expression plasmid, pRTLl1, contains a bacteriophage T5 promoter/lac operator element, which is controlled by lac repressor, supplied by the secondary plasmid, pBL1. Upon IPTG induction, a 90-kDa polyprotein is synthesised and subsequently proteolytically cleaved to reveal 64-kDa and 52-kDa polypeptides. Partial purification reveals that reverse transcriptase activity co-migrates with these two polypeptides.
Nature Genetics, 2000
Many pathological processes, including those causing allergies and autoimmune diseases, are assoc... more Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation 1 . Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes 2 . Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells.
Nature Genetics, 2000
Many pathological processes, including those causing allergies and autoimmune diseases, are assoc... more Many pathological processes, including those causing allergies and autoimmune diseases, are associated with the presence of specialized subsets of T helper cells at the site of inflammation 1 . Understanding the genetic program that controls the functional properties of T helper type 1 (Th1) versus T helper type 2 (Th2) cells may provide insight into the pathophysiology of inflammatory diseases. We compared the gene-expression profiles of human Th1 and Th2 cells using high-density oligonucleotide arrays with the capacity to display transcript levels of 6,000 human genes 2 . Here we analyse the data sets derived from five independent experiments using statistical algorithms. This approach resulted in the identification of 215 differentially expressed genes, encoding proteins involved in transcriptional regulation, apoptosis, proteolysis, and cell adhesion and migration. A subset of these genes was further upregulated by exposure of differentiated Th1 cells to interleukin-12 (IL-12), as confirmed by kinetic PCR analysis, indicating that IL-12 modulates the effector functions of Th1 cells in the absence of antigenic stimulation. Functional assays and in vivo expression of selected genes have validated the biological relevance of our study. Our results provide new insight into the transcriptional program controlling the functional diversity of subsets of T helper cells.
Nature Biotechnology, 1998
DNA Arrays, 2001
ABSTRACT Miniaturization and high-throughput parallel analysis are new concepts entering modern m... more ABSTRACT Miniaturization and high-throughput parallel analysis are new concepts entering modern molecular biology. An exciting example of such technology originally developed by physicists and applied to biology is the microchip. The semiconductor industry manufactures silica chips with increasing numbers of smaller and smaller features, which allows incredible numbers of operations within split seconds. The same principle of performing multiparallel operations on a miniaturized solid phase has led to the development of miniaturized arrays of several hundreds of thousands of DNA fragments on a small chip. Arraying of DNA or RNA samples onto nitrocellulose or nylon membranes is a very common analytical procedure in a molecular biology laboratory. DNA chips or microarrays are miniaturized versions of these classical filter-hybridization techniques. With great precision, thousands of DNA fragments or DNA oligonucleotides can be printed onto glass chips or microscope slides. In the case of oligonucleotides, they can also be synthesized directly on coated silica chips.
Nature Biotechnology, 1998
Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1979
1. Total RNA was extracted from human term placenta and mRNA purified by chromatography on oligo(... more 1. Total RNA was extracted from human term placenta and mRNA purified by chromatography on oligo(dT)-cellulose. The poly(A)-containing fraction stimulated amino acid incorporation 5- to 10-fold in the wheat germ cell-free system. Immunoprecipitation with an anti-lactogen serum indicated that 14-27% of the peptides synthesized in vitro contained antigenic determinants of this hormone. 2. Analysis of the [3H]leucine labelled product in the immunoprecipitate on sodium dodecyl sulfate-polyacrylamide gels revealed a complex mixture of polypeptides. Two heavily labelled bands (I and III) were seen corresponding in mobility with pre-lactogen (Mr = 25 000) and native lactogen (Mr = 22 200), each accounting for about 30% of the immunoprecipitable radioactivity. Two additional bands with an intermediate mobility were also observed. 3. Synthesis of the hormone was inhibited by 7-methylguanosine-5'-monophosphate suggesting the presence of a 7-methylguanosine 'cap' on the 5'-end of the mRNA for lactogen. 4. Peptide analysis of the cyanogen bromide cleavage products of band I, band III and authentic lactogen showed marked similarities in their primary structure. The precursor molecule, however, was lacking the N-terminal peptide present in authentic hormone indicating the presence of an extension of 25 amino acids at this side of the molecule. 5. The presence of one or several processing enzymes in the wheat germ cell-free system was indicated by the effect of Triton X-100. Low concentrations of this detergent (0.04%) while inhibiting the protein synthesizing activity for only 15%, completely abolished the precursor cleavage activity. Under these conditions only pre-lactogen was detected in the immunoprecipitate.
European Journal of Biochemistry, 1982
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1983
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1982
The amino acid sequence of component C1, the polypeptide specific for subunit F of prostatic bind... more The amino acid sequence of component C1, the polypeptide specific for subunit F of prostatic binding protein, the major secretory glycoprotein of the rat ventral prostate, has been determined. Its structure was established using the manual Edman degradation on the intact protein and on the most relevant fragments isolated from trypsin, chymotrypsin, thermolysin and Staphylococcus aureus protease digests of the 14C-labelled S-carboxamidornethylated component C1.Component C1 contains 88 amino acids corresponding to a molecular weight of 10246. It is an acidic polypeptide due to the presence of 17 acidic residues; its thrce cysteine residues are almost symmetrically distributed over the peptide chain. Highly polar regions are found in positions 17-27 and 37-47, while the C-terminal part of the molecule contains two hydrophobic segments
European Journal of Biochemistry, 1978
The prostatic binding protein, previously described in rat ventral prostate, was isolated. The pu... more The prostatic binding protein, previously described in rat ventral prostate, was isolated. The purified protein binds pregnenolone with an affinity of 1.2 × 106 M−1 and contains an average of 0.84 binding site per molecule. Its carbohydrate content is 3.2%. Its Mr, estimated by gel filtration, is 51000 but in the presence of 6 M guanidine hydrochloride or 0.1% dodecylsufate it dissociates into two subunits (S and F), which can be separated by polyacrylamide gel electrophoresis or by chromatography on hydroxyapatite. The Mr of these subunits is about 17000, when estimated by gel filtration in 6 M guanidine hydrochloride, or 19000 for subunit F and 20000 for subunit S, when measured by dodecylsulfate/polyacrylamide gel electrophoresis. Their isoelectric points, estimated by isoelectric focusing in 8 M urea, are 4.6 for subunit F and 4.9 for subunit S. Prostatic binding protein and both subunits have a very similar amino acid composition. Upon reduction of disulfide bridges each subunit dissociates further into two components: one of these components is the same in both subunits.
European Journal of Biochemistry - EUR J BIOCHEM, 2005
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1978
European Journal of Biochemistry, 1983
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1979
Polysomal RNA was extracted from human term placenta and total poly(A)-containing RNA purified by... more Polysomal RNA was extracted from human term placenta and total poly(A)-containing RNA purified by affinity chromatography on oligo(dT)-cellulose. Poly(A)-containing RNA constituted approximately 1.2 "/, of the total polysomal RNA and 8 % of this purified preparation was able to anneal with [3H
European Journal of Biochemistry, 1982
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 2005
residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues i... more residue 55 to 66 and from residue 77 up to 85. Remarkably four out of the six tyrosine residues in component C1 are located in the hydrophobic segment at the C terminus.
European Journal of Biochemistry, 1983