Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.3.1.28 (chloramphenicol acetyltransferase)
 5,100 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The gfa gene encodes glial fibrillary acidic protein, an intermediate filament protein found almost exclusively in astrocytes. Transient transfection studies with a chloramphenicol acetyltransferase reporter gene were used to identify regions of the gfa gene responsible for its expression. Three regions, A, B, and D, were found to be important. The D region is located near the basal promoter, while A and B are next to each other about 1500 bp further upstream. The regions contain several sequences homologous to binding sites of known transcription factors, and in addition, each contains an identical novel 10-bp motif. The A, B, and D regions act in a cell-specific manner; when joined to the SV 40 early promoter, they enhance transcription in the glial cell line U251, but not in the nonglial cell line HepG2. Consistent with this observation, the DNase I footprint produced in these regions by nuclear extract from U251 cells differs from that produced by an extract from HepG2 cells. The B region appears to be the most active of the three, as by itself it stimulates strong cell-specific transcription, whereas addition of the other two regions has little effect. When the B region is at its normal distance from the basal promoter, deletion of D severely reduces transcription, but when B is placed near the promoter, D is unimportant. This suggests that the D region may function primarily to promote interactions that bring B close to the promoter.
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PMID:Multiple interacting sites regulate astrocyte-specific transcription of the human gene for glial fibrillary acidic protein. 191 4

To develop a procedure for the transplantation of genetically modified brain primary cells, we transplanted cultured mouse cerebellar cells infected with recombinant retroviruses into the cerebella of adult mice and examined the expression of introduced gene-products in the host cerebellum after transplantation. After infection of cultured cerebellar cells with recombinant retroviruses harboring chloramphenicol acetyltransferase (CAT) gene, we selected only virus-infected cells for transplantation by culturing the cells in medium containing G418 for 3 weeks. CAT was continuously expressed in the cultured cerebellar cells during the 3-week incubation, but by immunoblotting analysis with antiglial fibrillar acidic protein (GFAP) or antineurofilament protein (NFP) antiserum the population of cultured cerebellar cells was found to change during the incubation. Immunocytochemical analyses using anti-CAT antiserum demonstrated that the transplanted cell mass containing CAT-positive cells was detectable in the cerebellum up to 3 weeks, but not 3 months after the transplantation of G418-selected cells into the cerebella of 7-week-old mice.
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PMID:Retrovirus-mediated gene transfer into mouse cerebellar primary culture and its application to the neural transplantation. 219 75

The functional induction of c-fos in the sodium butyrate-induced differentiation of F-98 glioma cells was studied. Fos protein level was increased by butyrate. In contrast, c-Jun protein was constitutively expressed and was not affected by butyrate. Gel-retardation assay indicates Fos as a component of the complex formed between the consensus oligonucleotide of the TPA (PMA, phorbol 12-myristate 13-acetate) response element (TRE) and nuclear extract prepared from butyrate-treated cells. Transfection studies showed that butyrate increased transcription from a multimeric TRE-driven reporter construct, and the effect was mimicked by transfecting cells with fos-expression plasmid. Furthermore, under conditions of c-fos over-expression, transactivation by butyrate was essentially abolished. These data suggest that Fos induction had a functional role in gene activation. Characterization of stable c-fos transfectants demonstrated that these cells displayed alterations in morphology, showed serum-dependent growth, had slower growth rates and grew to lower saturation densities than did untransfected F-98 cells or transfected cells that did not express c-fos. Immunofluorescent staining indicated that fos transfectants also had elevated glial fibrillary acidic protein ('GFAP') expression. Transfection of the c-fos promoter-chloramphenicol acetyltransferase fusion gene into F-98 cells revealed that activation of c-fos by butyrate was exerted at the promoter level, and sequences located within nucleotides -757 to -402 of the c-fos promoter were responsible for butyrate induction. Our data indicate that transcriptional activation of c-fos through its promoter by butyrate resulted in increased Fos protein expression. Transfection studies show that both c-fos and butyrate activate TRE-containing genes, and fos may be a downstream mediator of butyrate. Furthermore, expression of c-fos plays a major role in modulating the growth properties of F-98 cells.
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PMID:Analysis of c-fos expression in the butyrate-induced F-98 glioma cell differentiation. 786 28

A foreign gene coding the bacterial enzyme, chloramphenicol acetyltransferase (CAT), was introduced into a primary culture of the mouse cerebellar primordium by a retrovirus vector which harbors the neomycin-resistant gene. Following selection of the gene-transferred cells based on neomycin resistance, most of the selected cells expressed the CAT gene product as well as a marker for astrocytes, glial fibrillary acidic protein (GFAP), when examined immunocytochemically. These cells were transplanted into the adult mouse cerebellum, and the surviving cells were examined immunohistochemically by marking them with anti-CAT antibody. The distribution of CAT-immunopositive cells coincided with that of GFAP-immunopositive cells observed in serial sections of grafted sites at 10 days after transplantation. Some of the transplanted CAT-immunopositive cells extended processes and exhibited the morphological appearance of fibrous astrocytes. Migration of the genetically labeled cells into the host molecular layer was also observed and the morphological plasticity of the differentiated primary cells was shown according to the grafted sites. These results indicate that stable marking of cells for grafting can be accomplished by retrovirus-mediated introduction of a foreign gene into the primary culture, and that the fate and behavior of the labeled donor cells can be analyzed immunohistochemically following transplantation into neural tissue.
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PMID:Fate and behavior of genetically labeled cerebellar cells after transplantation into mouse cerebellum. 823 27

To understand astrocyte-specific transcription, we have been studying the human gfa gene. This gene encodes glial fibrillary acidic protein (GFAP), an intermediate filament protein expressed primarily in astrocytes. A survey of the gfa 5' flanking region showed it to contain several segments that contribute to expression of a chloramphenicol acetyltransferase reporter gene in transfected cells. The most active of these was the 124-bp B region, which spans bp -1612 to -1489. We have now used site-directed mutagenesis to analyze this region in greater detail, and show that the B region itself contains several important elements. The most crucial of these is a consensus AP-1 sequence, the binding site for the Fos and Jun families of transcription factors. The presence of members of both these families in the glial fibrillary acidic protein-expressing U251 cell line used for our transfection studies was verified by gel mobility-shift experiments. This is the first demonstration of the functioning of a specific transcription factor site for astrocytes, and provides a focus for future studies of glial fibrillary acidic protein regulation during development and reactive gliosis.
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PMID:Analysis of a segment of the human glial fibrillary acidic protein gene that directs astrocyte-specific transcription. 851 62