Transcription Initiation Site

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Akiyoshi Fukamizu - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Variation of the Human Angiotensinogen Core Promoter Element Located between the TATA Box and Transcription Initiation Site Affects Its Transcriptional Activity
    The Journal of biological chemistry, 1997
    Co-Authors: Kazuyuki Yanai, Tomoko Saito, Keiko Hirota, Hideyuki Kobayashi, Kazuo Murakami, Akiyoshi Fukamizu
    Abstract:

    Abstract Recent genetic studies indicate that several molecular variants discovered in angiotensinogen (AG), the precursor of vasoactive octapeptide angiotensin II, could potentially be responsible for inherited predisposition to human blood pressure variation. We have previously shown that a ubiquitously expressed nuclear factor, AGCF1, bound to AGCE1 (AG core promoterelement 1 including the core nucleotides,CTCGTG, CTC-type) located between the TATA box and Transcription Initiation Site (positions −25 to −1) is an authentic regulator of human AG Transcription. In the present study, we showed that AGCF1 has biologically and immunologically similar properties to those of a helix-loop-helix nuclear factor USF1 and examined the effects of two other naturally occurring molecular variants (ATCGTG, ATC-type and ATTGTG, ATT-type) found in the AGCE1 position on the human AG Transcriptional activity. Competitive gel-shift and transfection experiments demonstrated that the Transcriptional activity for the CTC- and ATC-type promoters was 2.5 times higher than that for the ATT-type through the alteration of AGCF1-binding affinity. These results suggest the possible involvement of USF1 as a component in AGCF1 formation and the potential importance of AGCE1 variation in blood pressure regulation through human AG expression.

  • a cis acting dna element located between tata box and Transcription Initiation Site is critical in response to regulatory sequences in human angiotensinogen gene
    Journal of Biological Chemistry, 1996
    Co-Authors: Kazuyuki Yanai, Kazuo Murakami, Yutaka Nibu, Akiyoshi Fukamizu
    Abstract:

    Abstract The promoter of the human angiotensinogen (hAG) gene functioned in its own core promoter context but not when replaced with simian virus 40 (SV40) core promoter, suggesting the presence of a Transcriptionally important cis-acting sequence. Electrophoretic mobility shift assays demonstrated that a ubiquitously expressed nuclear factor, AGCF1, bound to AGCE1 (hore promoter lement 1; positions −25 to −1) located between the TATA box and Transcription Initiation Site. Substitution mutation in AGCE1 which disrupted AGCF1 binding affected the promoter activity more severely than a nonsense mutation of the hAG TATA sequences did. When AGCE1 was placed at the downstream of SV40 core promoter, the responsiveness to hAG upstream region was significantly restored. Furthermore, mutation and in vivo competition experiments suggested that AGCF1 acts as a critical regulator of hAG Transcription by mediating the activity of the hAG upstream and downstream enhancer elements. DNase I footprinting and UV cross-linking analyses showed that AGCF1 with apparent molecular masses of 31, 33, and 43 kDa as the components protected the region from −26 to −9 which partially overlapped with the TATA box consensus sequences. These findings indicate that AGCE1 in addition to the TATA box plays a key role in mediating the hAG regulatory elements.

Jennifer Wang - One of the best experts on this subject based on the ideXlab platform.

  • An Embryonic Alternative Transcription Initiation Site and the 5′-Upstream Structure of Mouse Cardiac Troponin T Gene
    Biochemical and biophysical research communications, 1995
    Co-Authors: Jian Ping Jin, Jinyi Zhang, Jennifer Wang
    Abstract:

    Abstract By specific antiboby screening of a λZAPII expression library, we identified a class of cDNAs derived from mouse cardiac troponin T (TnT) mRNA with long 5′-ends containing the -30 TATA sequence of the promoter. These long mRNAs indicated the presence of a novel upstream Transcription Initiation Site in the cardiac TnT gene. Primer extension mapping of the mouse cardiac TnT mRNA confirmed an alternative Transcription Initiation Site 55-base pairs Initiation Site. The upstream Initiation Site was found significantly more active in the embryonic/neonatal hearts. Genomic cloning and sequencing further revealed an 1.5-kb 5′-upstream sequence of the mouse cardiac TnT gene and relationships between the promoter regulatory elements and the alternative Transcription Initiation Sites.

  • an embryonic alternative Transcription Initiation Site and the 5 upstream structure of mouse cardiac troponin t gene
    Biochemical and Biophysical Research Communications, 1995
    Co-Authors: Jian Ping Jin, Jinyi Zhang, Jennifer Wang
    Abstract:

    Abstract By specific antiboby screening of a λZAPII expression library, we identified a class of cDNAs derived from mouse cardiac troponin T (TnT) mRNA with long 5′-ends containing the -30 TATA sequence of the promoter. These long mRNAs indicated the presence of a novel upstream Transcription Initiation Site in the cardiac TnT gene. Primer extension mapping of the mouse cardiac TnT mRNA confirmed an alternative Transcription Initiation Site 55-base pairs Initiation Site. The upstream Initiation Site was found significantly more active in the embryonic/neonatal hearts. Genomic cloning and sequencing further revealed an 1.5-kb 5′-upstream sequence of the mouse cardiac TnT gene and relationships between the promoter regulatory elements and the alternative Transcription Initiation Sites.

Kazuyuki Yanai - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Variation of the Human Angiotensinogen Core Promoter Element Located between the TATA Box and Transcription Initiation Site Affects Its Transcriptional Activity
    The Journal of biological chemistry, 1997
    Co-Authors: Kazuyuki Yanai, Tomoko Saito, Keiko Hirota, Hideyuki Kobayashi, Kazuo Murakami, Akiyoshi Fukamizu
    Abstract:

    Abstract Recent genetic studies indicate that several molecular variants discovered in angiotensinogen (AG), the precursor of vasoactive octapeptide angiotensin II, could potentially be responsible for inherited predisposition to human blood pressure variation. We have previously shown that a ubiquitously expressed nuclear factor, AGCF1, bound to AGCE1 (AG core promoterelement 1 including the core nucleotides,CTCGTG, CTC-type) located between the TATA box and Transcription Initiation Site (positions −25 to −1) is an authentic regulator of human AG Transcription. In the present study, we showed that AGCF1 has biologically and immunologically similar properties to those of a helix-loop-helix nuclear factor USF1 and examined the effects of two other naturally occurring molecular variants (ATCGTG, ATC-type and ATTGTG, ATT-type) found in the AGCE1 position on the human AG Transcriptional activity. Competitive gel-shift and transfection experiments demonstrated that the Transcriptional activity for the CTC- and ATC-type promoters was 2.5 times higher than that for the ATT-type through the alteration of AGCF1-binding affinity. These results suggest the possible involvement of USF1 as a component in AGCF1 formation and the potential importance of AGCE1 variation in blood pressure regulation through human AG expression.

  • a cis acting dna element located between tata box and Transcription Initiation Site is critical in response to regulatory sequences in human angiotensinogen gene
    Journal of Biological Chemistry, 1996
    Co-Authors: Kazuyuki Yanai, Kazuo Murakami, Yutaka Nibu, Akiyoshi Fukamizu
    Abstract:

    Abstract The promoter of the human angiotensinogen (hAG) gene functioned in its own core promoter context but not when replaced with simian virus 40 (SV40) core promoter, suggesting the presence of a Transcriptionally important cis-acting sequence. Electrophoretic mobility shift assays demonstrated that a ubiquitously expressed nuclear factor, AGCF1, bound to AGCE1 (hore promoter lement 1; positions −25 to −1) located between the TATA box and Transcription Initiation Site. Substitution mutation in AGCE1 which disrupted AGCF1 binding affected the promoter activity more severely than a nonsense mutation of the hAG TATA sequences did. When AGCE1 was placed at the downstream of SV40 core promoter, the responsiveness to hAG upstream region was significantly restored. Furthermore, mutation and in vivo competition experiments suggested that AGCF1 acts as a critical regulator of hAG Transcription by mediating the activity of the hAG upstream and downstream enhancer elements. DNase I footprinting and UV cross-linking analyses showed that AGCF1 with apparent molecular masses of 31, 33, and 43 kDa as the components protected the region from −26 to −9 which partially overlapped with the TATA box consensus sequences. These findings indicate that AGCE1 in addition to the TATA box plays a key role in mediating the hAG regulatory elements.

Antoine R Stuitje - One of the best experts on this subject based on the ideXlab platform.

  • sequences surrounding the Transcription Initiation Site of the arabidopsis enoyl acyl carrier protein reductase gene control seed expression in transgenic tobacco
    Plant Molecular Biology, 1999
    Co-Authors: Gertjan De Boer, Christa Testerink, Gerlof Pielage, John H J Nijkamp, Antoine R Stuitje
    Abstract:

    The NADH-specific enoyl-acyl carrier protein (ACP) reductase, which catalyses the last reducing step during the fatty acid biosynthesis cycle, is encoded in Arabidopsis thaliana encoded by a single housekeeping gene (ENR-A) which is differentially expressed during plant development. To identify elements involved in its tissue-specific Transcriptional control, a fragment comprising the 1470 bp region directly upstream of the ATG start codon of the ENR-A gene was fused to the uidA (GUS) reporter gene and analysed in transgenic Nicotiana tabacum plants. GUS activity found during development of the transgenic plants was similar to endogenous ENR protein levels found in both tobacco and Arabidopsis plants, except for developing flowers. In floral tissue the promoter fragment showed very little activity in contrast to the relatively high level of endogenous ENR expression. Successive deletions from the 5′ and 3′ regions of the promoter fragment revealed the presence of at least three elements which control GUS expression in different stages of development in the transgenic tobacco plants. First, expression in young developing leaves required both the presence of sequences between −329 to −201 relative to the Transcription start and part of the untranslated leader comprising the first intron. Second, root-specific GUS expression was still observed after deletion of the 5′-upstream sequences up to 19 bp of the Transcription Initiation Site. Further, the additional removal of the intron from the untranslated leader increased root-specific expression by ca. 4- to 5-fold. Third, high expression in seeds was still observed with the minimal upstream promoter segment of 19 bp. This seed expression level was found to be independent of the presence or absence of the intron in the untranslated leader. Finally, 3′ deletion of the leader sequence up to 17 bp of the Transcription start greatly impaired GUS activity during all stages of plant development, suggesting that the deleted sequence of the leader either functions as an enhancer for Transcription Initiation or stabilizes the mRNA.

P.i. Schrier - One of the best experts on this subject based on the ideXlab platform.

  • HLA-B locus-specific downregulation in human melanoma requires enhancer A as well as a sequence element located downstream of the Transcription Initiation Site.
    Immunogenetics, 2000
    Co-Authors: Marieke Griffioen, Ilse J.m. Ouwerkerk, Veronique Harten, P.i. Schrier
    Abstract:

    Human MHC class I molecules are encoded by three different loci (HLA-A, -B, and -C), which are regulated at the Transcriptional level through several conserved cis-acting promoter elements. The presence of locus-specific residues throughout the entire promoter region strongly suggests that the various HLA class I loci are differentially regulated. To identify regulatory sequences involved in locus-specific HLA class I gene Transcription, a series of truncated HLA-A2 and HLA-B7 promoter-reporter constructs were transfected into melanoma cell lines expressing high and low levels of endogenous HLA-B, but comparable levels of HLA-A. These experiments showed that differential regulation of HLA-B expression in melanoma cell lines is mediated by a previously unidentified co-operative action of enhancer A, located 175 bp upstream of the Transcription Initiation Site (+1), and a specific region of 20 nucleotides located at +13 to +33 bp downstream of the Transcription Initiation Site. Furthermore, we demonstrated binding of Transcription factor Yin Yang 1 to the HLA-A +13/+33 bp region, but not to the equivalent HLA-B region. Based on these results, we present a model suggesting that YY1 displaces either activating or repressing Transcription factors, thereby making the HLA-A gene resistant to differential regulation.

  • HLA-B down-regulation in human melanoma is mediated by sequences located downstream of the Transcription-Initiation Site.
    International journal of cancer, 1999
    Co-Authors: Marieke Griffioen, Ilse J.m. Ouwerkerk, Veronique Harten, P.i. Schrier
    Abstract:

    Major histocompatibility complex (MHC, HLA in humans) class I molecules play an important role in cellular immunology by presenting viral, tumor-associated or minor histocompatibility antigen-derived peptides to T cells. Tumor cells frequently fail to express one or more of the different MHC class I loci (HLA-A, -B and -C), thereby avoiding elimination by T cells. In primary human melanomas as well as melanoma cell lines, HLA class I expression is frequently down-regulated in a B locus-specific manner. The HLA class I promoter contains a number of cis-regulatory elements located upstream of the Transcription-Initiation Site, among them enhancer A and an interferon-stimulated response element. In the present study, we show that novel sequences located 13 to 33 bp downstream of the Transcription-Initiation Site mediate HLA-B locus-specific down-regulation in human melanoma cell lines. Furthermore, involvement of the +13 to +33-bp region in HLA-B locus-specific down-regulation in vivo is supported by in vitro experiments showing locus-specific binding of protein complexes to the +13 to +33-bp region. Int. J. Cancer 80:573–580, 1999. © 1999 Wiley-Liss, Inc.