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E Gizang-Ginsberg

Publications and source records attributed to E Gizang-Ginsberg.

10 recordsLinked to original sources
PubMed1995

Transcriptional activators differ in their responses to overexpression of TATA-box-binding protein.

We investigated how overexpression of human TATA-box-binding protein (TBP) affects the action of estrogen receptor (ER) and compared the response with that of other activators. When ER activates a simple promoter, consisting of a response element and either the collagenase or tk TATA box, TBP overexpression potentiates transcription. TBP potentiates only estrogen-induced and not basal transcription and does so independent of spacing between response element and TATA box. TBP overexpression also reduces autoinhibition by overexpressed ER, suggesting that one target of the autoinhibition may be TBP itself. Both AF-1 and AF-2 domains of ER are potentiated by TBP, and each domain binds TBP in vitro. Like ER, chimeric GAL4/VP16 and GAL4/Tat activators are also potentiated by TBP, as is the synergistic activation by ER and GAL4/VP16 on a complex promoter. Unlike ER, GAL4/Sp1 and GAL4/NF-I become less potent when TBP is overexpressed. Furthermore, synergy between ER and Sp1 or between ER and NF-I, whether these are supplied by transfected GAL4 fusions or by the endogenous genes, is inhibited by TBP overexpression. Thus, ER resembles VP16 in response to TBP overexpression and is different from Sp1 and NF-I, which predominate over ER in setting the response on complex promoters.

Animals↗
PubMed1994

Fos family members successively occupy the tyrosine hydroxylase gene AP-1 site after nerve growth factor or epidermal growth factor stimulation and can repress transcription.

Nerve growth factor induces the neuronal-like differentiation of PC12 cells, and epidermal growth factor promotes PC12 viability and is weakly mitogenic. Despite these differences, both growth factors induce indistinguishable patterns of transient delayed transcription of the tyrosine hydroxylase (TH) gene and the expression of proteins encoded by Fos gene family members. Thus, TH expression is sensitive to signaling pathways common to these two growth factors. We show that c-fos and fosB successively occupy an AP-1 site-like element of the TH promoter after nerve growth factor treatment. Furthermore, under conditions of transient transfection, Fos family proteins may synergize with c-jun to transrepress TH gene transcription through the TH-fat-specific element. We show that the target of repression is the AP-1 site-like element that lies within the TH-fat-specific element. We demonstrate that this site is also a major positive acting site for TH control. These results suggest a model in which the long term effect of c-fos family protein expression is to limit the expression of the TH gene. We consider the novel properties of this element in providing temporal and cell type-specific regulation of TH transcription.

Animals↗
PubMed1992

Nerve growth factor-induced derepression of peripherin gene expression is associated with alterations in proteins binding to a negative regulatory element.

The peripherin gene, which encodes a neuronal-specific intermediate filament protein, is transcriptionally induced with a late time course when nerve growth factor (NGF) stimulates PC12 cells to differentiate into neurons. We have studied its transcriptional regulation in order to better understand the neuronal-specific end steps of the signal transduction pathway of NGF. By 5' deletion mapping of the peripherin promoter, we have localized two positive regulatory elements necessary for full induction by NGF: a distal positive element and a proximal constitutive element within 111 bp of the transcriptional start site. In addition, there is a negative regulatory element (NRE; -179 to -111), the deletion of which results in elevated basal expression of the gene. Methylation interference footprinting of the NRE defined a unique sequence, GGCAGGGCGCC, as the binding site for proteins present in nuclear extracts from both undifferentiated and differentiated PC12 cells. However, DNA mobility shift assays using an oligonucleotide probe containing the footprinted sequence demonstrate a prominent retarded complex in extracts from undifferentiated PC12 cells which migrates with slower mobility than do the complexes produced by using differentiated PC12 cell extract. Transfection experiments using peripherin-chloramphenicol acetyltransferase constructs in which the footprinted sequence has been mutated confirm that the NRE has a functional, though not exclusive, role in repressing peripherin expression in undifferentiated and nonneuronal cells. We propose a two-step model of activation of peripherin by NGF in which dissociation of a repressor from the protein complex at the NRE, coupled with a positive signal from the distal positive element, results in depression of the gene.

Animals↗
PubMed1990

Nerve growth factor regulates tyrosine hydroxylase gene transcription through a nucleoprotein complex that contains c-Fos.

We have studied nerve growth factor (NGF) regulation of the expression of the tyrosine hydroxylase (TH) gene in PC12 cells. The TH gene encodes the initial and rate-limiting enzyme of the catecholamine biosynthetic pathway. We show that the TH gene is transiently transcriptionally induced by a mechanism reliant on new protein synthesis during 1-2 hr of NGF stimulation, a time following the induction of the c-fos gene at 15 min post-NGF treatment. A potential regulatory sequence located within the TH gene promoter, the TH-FSE, shares homology to a known regulatory element, the fat-specific element (FSE), which is found upstream from genes activated during adipocyte differentiation and binds the Fos-Jun transcription factor complex. We show that the TH-FSE DNA sequence elevates the basal level of transcription from the rat TH promoter and is required for NGF inducibility. This DNA element binds authentic Fos-Jun products produced abundance during NGF stimulation and by in vitro translation. We demonstrate further that the TH-FSE can bind proteins present in PC12 nuclear extracts in a sequence-specific manner. The DNA/nucleoprotein complex that forms increases in abundance during NGF stimulation and reaches a maximum level at 4 hr of treatment. Antibody inhibition studies utilizing an anti-Fos antibody indicate that Fos and/or Fos-related antigen(s) associate with the TH-FSE and suggest that the Fos protein family contributes to the regulation of TH in vivo. These results support a model in which NGF-induced immediate early genes, including c-Fos, contribute to the regulation of delayed early genes such as TH and thereby control neuronal differentiation.

Animals↗
PubMed1987

Differential expression of the mouse homeobox-containing gene Hox-1.4 during male germ cell differentiation and embryonic development.

Hox-1.4 is a mouse homeobox-containing gene (initially identified as HBT-1), whose expression appears to be testis-specific in the adult animal. Examination of Hox-1.4 transcripts in RNA from testes of mutant mice deficient in germ cells confirms that Hox-1.4 expression within the testis is germ cell-specific. Enriched populations of spermatogenic cells were used to localize the expression of Hox-1.4 specifically to germ cells that have entered into and progressed beyond the meiotic prophase stage of differentiation and to demonstrate the presence of two different size Hox-1.4 transcripts. Examination of RNA from teratocarcinoma cell cultures and mouse embryos at 10.5-16.5 days of gestation demonstrated the presence of several Hox-1.4 transcripts, which are larger than those present in germ cells. In the midgestation fetus, Hox-1.4 expression is most abundant in the spinal cord.

Alleles↗
PubMed1987

Expression of the proopiomelanocortin gene is developmentally regulated and affected by germ cells in the male mouse reproductive system.

Proopiomelanocortin (POMC), a major pituitary product, is also present in the adult mouse testis. We have shown previously that POMC mRNAs are most abundant in a subpopulation of Leydig cells associated with tubules in specific stages of the cycle of the seminiferous epithelium. In the present study, we examined the expression of the gene encoding POMC during testicular development and in other tissues of the male reproductive system. We also analyzed the effects of cellular interactions on POMC gene expression in the testis. Blot-hybridization analysis revealed that POMC transcripts of approximately equal to 800 nucleotides were present in enriched populations of meiotic prophase spermatocytes and in caput epididymis but were absent in cauda epididymis and vas deferens. POMC transcripts were present in fetal testis (day 17 of gestation to newborn), could not be detected in prepuberal testis (days 7-8 postpartum), but reappeared in the adult testis. No difference in the size or abundance of POMC transcripts was seen in testes from mouse mutant strains in which spermatogenesis is arrested in early spermiogenesis. In contrast, POMC transcripts were virtually undetectable in testes that are devoid of germ cells. These results emphasize the importance of interactions between germ cells and interstitial cells and the regulation of the POMC gene in the mammalian testis.

Animals↗
PubMed1986

Isolation of a mouse cDNA coding for a developmentally regulated, testis-specific transcript containing homeo box homology.

A clone, pHBT-1, containing sequences homologous to Drosophila homeo boxes has been isolated from a mouse testis cDNA library. The sequence is 80% homologous at the DNA level and 88% homologous at the amino acid level to the homeo box sequence of the Antennapedia gene of Drosophila. Sequences flanking the 3' end of the homeo box are highly diverged from other murine homeo box-containing genes characterized to date. RNA blot hybridization analysis of mouse testis poly(A)+ RNA revealed transcripts of approximately 1.4 kb in length. Within the limits of sensitivity of detection of Northern blot analysis, no transcripts were seen in any of the adult somatic tissues examined. Other tissues that contain stem cells, namely those of the hemopoietic system, also lacked detectable amounts of HBT-1 transcripts. HBT-1 transcripts were limited to male germ cell-containing tissues, since RNAs from juvenile and adult ovaries did not contain detectable amounts of the 1.4-kb transcripts. Expression of the HBT-1 gene was not detected in embryonic testes, nor in tests of neonatal animals which contain germ cells up to the Type B stage of spermatogonial development. A role for the expression of the HBT-1 gene in the meiotic stages of male germ cell differentiation is postulated.

Animals↗
PubMed1985

Separation of mouse testis cells on a Celsep (TM) apparatus and their usefulness as a source of high molecular weight DNA or RNA.

The use of a self-contained unit-gravity cell separation apparatus for separation of populations of mouse testicular cells is described. The apparatus, a Celsep (TM), maximizes the unit area over which sedimentation occurs, reduces the amount of separation medium employed, and is quite reproducible. Cells thus isolated have been good sources for isolation of DNA, and notably, high molecular weight RNA.

Animals↗
PubMed1985

Localization of mRNAs in mouse testes by in situ hybridization: distribution of alpha-tubulin and developmental stage specificity of pro-opiomelanocortin transcripts.

Gene expression of mouse testicular germ cells and surrounding somatic cells during spermatogenesis was examined by RNA:cDNA hybridization in situ and concomitant Northern and dot blot analysis. Particular attention was paid to obtaining fixation and hybridization procedures for use with mouse testes to accomplish sensitive and precise localization with the in situ technique. alpha-Tubulin mRNAs were localized in virtually all testis cell types. In elongating spermatids, a unique labeling pattern was visualized; possibly corresponding to the position of the asymmetrically displaced cytoplasm. Pro-opiomelanocortin (POMC) transcripts in the adult mouse testis detected by Northern blot analysis were shown to be of a smaller size (approximately 600-800 nt) than the pituitary POMC transcripts, similar to what has been reported recently for POMC expression in the rat testis and mouse Leydig cell lines. Localization of POMC mRNAs by in situ hybridization was primarily to Leydig cells, although some labeling of spermatogonia and spermatocytes within the seminiferous epithelium was detected. A cellular or developmental specificity of the pattern of POMC localization was observed: obvious labeling of Leydig cells was limited to interstitial regions which were surrounded completely or in part by tubules in stages IX to XII of the cycle of the seminiferous epithelium.

Animals↗