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O Bakker

Publications and source records attributed to O Bakker.

44 records · Page 3Linked to original sources

CAAT/enhancer binding protein is able to bind to ATF/CRE elements.

The transcription factor C/EBP is known to be able to bind to two different classes of sites, a CAAT-box and one present in a number of viral enhancers. In this paper we show using band-shift assays, methylation interference and footprinting that C/EBP is also able to bind with high affinity to ATF/CRE sites. Competition with mutant ATF sites and methylation interference indicate that C/EBP may be able to bind to the ATF/CRE sites by virtue of their homology to the enhancer core elements. Furthermore, we show that C/EBP is able to direct transcription from this site in transient transfection experiments and that mutations in the ATF binding site that impair DNA binding also effect the ability of C/EBP to stimulate transcription.

Activating Transcription Factors↗

The progesterone receptor can regulate transcription in the absence of a functional TATA box element.

We have investigated the importance of the TATA box element in the induction of transcription by the progesterone receptor. Transcription was analyzed from promoters containing a steroid response element upstream of a wild-type or mutated TATA box. Mutation of the TATA box resulted in a loss of correctly initiated transcripts and abolished binding of TATA factor to the TATA box in vitro but did not inhibit transcriptional activation by the progesterone receptor. Thus we conclude that the receptor is able to stimulate the rate of transcription in the absence of a functional TATA box.

Animals↗

Tissue-specific differences in the binding of nuclear proteins to a CCAAT motif in the promoter of the androgen-regulated C3 gene.

The expression of the gene for the C3 polypeptide is confined to the ventral prostate of the male rat and is regulated by androgens. To study the mechanism of this tissue-specific and hormone-dependent regulation we have used DNase-I footprinting and band-shift assays to locate binding sites for nuclear proteins isolated from different tissues. In this paper we present evidence that there are tissue-specific differences in the nuclear proteins that are able to bind to a CCAAT motif and a nuclear factor-I consensus that are present in the promoter of the rat C3 gene. Using competition assays and heat denaturation we show that the CAAT box/enhancer binding protein itself may be one of the transcription factors involved.

Androgen-Binding Protein↗

Turnover products of the apo very low density lipoprotein II messenger RNA from chicken liver.

The mature apo Very Low Density Lipoprotein II (apo VLDLII) mRNA appears in chicken liver within a few hours after estrogen administration. Apart from this mRNA species, shorter RNA molecules hybridizing to apo VLDLII sequences have been detected in rooster liver upon estrogen stimulation. These molecules are present in the non-polyadenylated fraction of the total cellular- and polysomal RNA. Northern blotting and electron microscopy of R-loops were employed to show that these shorter RNA molecules are truncated at their 3'-end. The 3'-termini were further characterized by nuclease S1 analyses, and are located predominantly in the 3' untranslated region of the mRNA. Using a secondary structure model (Shelness and Williams, J. Biol. Chem. 260, 8637-8646, 1985), we show that the 3' termini map mainly in unpaired regions of the structure.

Animals↗

Protein-DNA interactions in vitro with 5'-flanking DNA fragments from the chicken vitellogenin gene.

The expression of the vitellogenin gene in the liver of oviparous animals is under strict control of estrogen. We have studied the interaction of proteins extracted from nuclei of different estrogen responsive tissues with two fragments (-728 to -470 and -625 to -470) of the upstream region of the chicken vitellogenin gene, using the gel-retardation technique. We found a complex pattern of retarded bands using nuclear extracts from laying hen liver, rooster liver and MCF-7 cells. The patterns observed display differences in the position and intensities of some of the bands, depending on the source of the extract used. The possible significance of these findings will be discussed.

Animals↗

Estrogen-inducible binding of a nuclear factor to the vitellogenin upstream region.

The estrogen-dependent binding of a protein to the upstream region of the chicken vitellogenin gene was detected by using in vivo dimethyl sulfate, genomic DNase I, and in vitro exonuclease III footprinting. The site is located between base pairs -848 and -824, and its sequence resembles that of the nuclear factor I binding site. The results suggest that a nuclear factor binding to this site is involved in the regulation of the vitellogenin gene.

Animals↗

Differential estrogen responsiveness of the vitellogenin and apo very low density lipoprotein II genes in the rooster liver.

The primary transcript of the chicken apo Very Low Density Lipoprotein II (apoVLDL-II) gene is formed almost immediately after a first estrogen administration, contrary to the appearance of the vitellogenin primary transcript which has a lag of at least 4 h. However, after a second estrogen administration the vitellogenin gene transcription shows no detectable lag (memory effect). After estrogen withdrawal, the primary transcripts of both genes rapidly decline to undetectably low levels. In the presence of estrogen, the half-lives of the mRNAs of apoVLDL-II and vitellogenin are 15 and at least 70 h, respectively, whereas in the absence of hormone they are only 3.5 and 5.5 h, respectively. Thus estrogen not only controls the transcription of both genes, but also the turnover of their mRNAs. In short, there appears to be a quantitative difference in the response of both genes.

Animals↗

Induction of ornithine decarboxylase and augmentation of tyrosine aminotransferase activity by N-hydroxy-2-acetylaminofluorene and 2-acetylaminofluorene in rat liver. Influence of sex, retinylacetate, indomethacin, and pentachlorophenol.

IP injection in rats of 2-acetylaminofluorene (AAF) or N-hydroxy-2-acetylaminofluorene (N-OH-AAF) resulted in a transient increase of hepatic ornithine decarboxylase (ODC) and tyrosine aminotransferase (TAT) activity. Maximal activity of ODC was observed 4 hr and of TAT 3 hr after administration of either AAF or N-OH-AAF. A lag-time of 2 hr preceded the increase of ODC and TAT activity. N-OH-AAF dependent ODC induction displayed an almost linear dose-response in the dose range up to 94.1 mumol/kg bw (body weight) when the ODC activity was measured at its maximum 4 hr after administration. Elevation of the dose N-OH-AAF to 126 mol/kg bw resulted in a lower ODC induction. Administration of doses AAF to 31.4 mumol did not change ODC activity. At doses up to 126 mumol/kg bw ODC induction increased linear. TAT induction increased linear in the dose range 15.7-94.1 mumol N-OH-AAF and 31.4-94.1 mumol AAF/kg. Lowering the dose of AAF did not result in a lower ODC or TAT activity. Judged by the effects of actinomycin D or cycloheximide administered 1 hr prior to AAF or N-OH-AAF, the in vivo induction of rat liver ODC activity by AAF and N-OH-AAF appeared to be under transcriptional control, whereas augmentation of TAT activity under influence of AAF or N-OH-AAF appeared the result of (post) translational events. Induction of ODC by AAF or N-OH-AAF was not significantly changed by indomethacin, was slightly increased by pentachlorophenol (PCP) and was synergistically enhanced by retinylacetate (RA). The increase of TAT activity was stimulated by PCP and RA. The effect of PCP indicates that N-sulfonoxy-2-acetylaminofluorene is most probably not involved in the induction of ODC. AAF appeared more effective hepatic ODC inducer in females than males and moreover more effective than N-OH-AAF in females. N-OH-AAF had stronger ODC inducing capacity in males than females. Similar observations were made with respect to TAT activity. When induction of ODC is indicative for a tumor promoting property then the data presented here suggest that tumor promotion of the complete carcinogens AAF and N-OH-AAF is not mediated by N-O-sulfation; this might be due to other metabolic conversions.

2-Acetylaminofluorene↗