Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.30.1 (S1 nuclease)
 3,660 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The human insulin-like growth factor II (IGF-II) gene contains four promoters (P1, P2, P3 and P4). In order to determine the mechanism by which the P4 promoter is controlled, the human IGF-II P4 promoter was analyzed in cell lines. DNA sequence analysis of the human IGF-II P4 promoter gene showed that the P4 promoter region contains a TATA-like sequence and several G+C rich regions which are essential for transcription. Analysis of the transcription initiation site by S1 nuclease mapping revealed two transcription start sites; both are located immediately behind TATA-like sequence. To determine the location of sites that may be important for the function of the human IGF-II P4 promoter, we constructed chimeric genes of the human IGF-II P4 promoter fused to the coding region for chloramphenicol acetyltransferase (CAT). These constructs were transfected into HepG2, PLC/PRF/5, G401 and A549 cells, and were examined for CAT activity. All transfected cells showed a similar profile of CAT activity. Sequences responsible for putative enhancer and silencer regions were identified and the 5' flanking sequences of the human IGF-II P4 promoter contain negative regulatory regions (-213 to -174). The 53-base pair fragment located between 111 and 59 base pairs upstream of the start site contains positive regulatory activity. Gel mobility shift assay showed that Sp1 and another proteins might be involved in positive regulation of the human IGF-II P4 promoter.
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PMID:Characterization of the P4 promoter region of the human insulin-like growth factor II gene. 840 33

The gene encoding the human endothelin-B receptor (hET-BR) has been isolated, and its structural organization and chromosomal assignment have been determined. Southern blot analysis demonstrated a single copy of the hET-BR gene in the human genome. The hET-BR gene spans 24 kilobases and consists of seven exons and six introns. The size range for exons is 109-2855 base pairs, although that for introns is 0.2-14.5 kilobases. Every intron occurs near the border of the putative transmembrane domain in the coding region. The major transcription initiation sites were mapped to the positions 258 and 229 base pairs upstream of the ATG initiation codon by primer extension and nuclease S1 protection experiments. The 5'-flanking region of the hET-BR gene lacks conventional TATA and CCAAT boxes but contains a sequence of potential Sp1 binding sites upstream of the transcription initiation sites. There are some canonical consensus sequences of cis-elements including GATA motif, acute phase reactant regulatory element, and E box. Using human-rodent somatic hybrid cell lines, the hET-BR gene was assigned to human chromosome 13. The present study will lead to a better understanding of the mechanism for the transcriptional regulation of the hET-BR gene and will give a clue as to how to search for possible genetic disorders of hET-BR.
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PMID:The human endothelin-B receptor gene. Structural organization and chromosomal assignment. 842 23

The c-erbA alpha gene encodes the alpha type thyroid hormone receptor. This gene is expressed in various types of cells, its expression being relatively high in the central nervous system. A genomic clone that contains the 5'-terminal portion of the human c-erbA alpha gene was isolated. The 615 base pair (bp) 5'-flanking sequence of the c-erbA alpha gene showed promoter activity when placed upstream of the bacterial chloramphenicol acetyltransferase gene and transfected into HeLa cells. Nine transcriptional initiation sites were detected within this sequence by S1 nuclease protection analysis. DNA sequence analysis showed that the promoter region contains ten putative binding sites for transcriptional factor Sp1 in the GC rich region (86%). Three putative cAMP responsive elements (CRE) and one putative TPA responsive element (TRE) were identified upstream of the GC rich region. The c-erbA alpha promoter sequence also contains a putative binding site for the Krox-20 transcriptional factor, which is thought to play a role in early development of the mouse central nervous system.
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PMID:Molecular cloning and characterization of the promoter region of the human c-erbA alpha gene. 846 21

In this study, the minimal promoter requirements of the TATA-less human androgen receptor (hAR) gene promoter are described. The hAR promoter is characterized by a short GC-box (-59/-32) and a long homopurine stretch (-117/-60). Two major transcription initiation sites, AR transcription initiation site I (AR-TIS I, (+1/2/3)) and AR transcription initiation site II (AR-TIS II, (+12/13)) are located in a 13-base pair region (Faber, P. W., van Rooij, H. C. J., van der Korput, J. A. G. M., Baarends, W. M., Brinkmann, A. O., Grootegoed, J. A., and Trapman, J. (1991) J. Biol. Chem. 266, 10743-10749). Transient transfection of COS cells with hAR promoter deletion and mutant constructs, followed by RNA isolation and S1 nuclease protection analysis showed that the process of transcription initiation through AR-TIS I and AR-TIS II is regulated by different promoter sequences. The GC-box directed initiation from AR-TIS II but did not affect AR-TIS I utilization, which is dependent upon sequences between positions -5 and +57. Band shift analysis identified the transcription factor Sp1 as the protein interacting with the GC-box. A single Sp1 binding sequence was found to be present in the GC-box. Footprint analysis confirmed the interaction of Sp1 with this sequence. The differential initiation through AR-TIS I and AR-TIS II was substantiated by the introduction of point mutations in the Sp1 binding sequence: only mutations that specifically abolished Sp1 binding interfered with AR-TIS II utilization, but all mutations left AR-TIS I initiation intact.
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PMID:Two different, overlapping pathways of transcription initiation are active on the TATA-less human androgen receptor promoter. The role of Sp1. 848 25

The activity of the apical membrane Na+/H+ exchanger NHE3 isoform of renal or intestinal epithelial cells is chronically regulated by a wide variety of stimuli, including acidosis, cAMP, glucocorticoids, and thyroid hormone. To understand the molecular mechanisms responsible for long term regulation of this cation transporter, we have isolated and determined the structure of this gene from a rat genomic library. The Nh3 gene spans > 40 kilobases and contains 17 exons that are flanked by typical splice donor and acceptor sequences at the exon-intron boundaries. The transcription initiation site was mapped by S1 nuclease protection analyses of mRNA from rat kidney and intestine. Multiple start sites were clustered between nucleotides -100 and -96 relative to the translation initiation codon. An atypical TATA-box and CCAAT-box are centered 30 and 147 nucleotides, respectively, upstream of the predominant transcription initiation site. Sequence analysis of approximately 1.4 kilobases of the 5'-flanking promoter region also revealed the presence of other putative cis-acting elements recognized by various transcription factors (e.g. AP-1, AP-2, C/EBP, NF-I, OCT-1/OTF-1, PEA3, Sp1, glucocorticoid, and thyroid hormone receptors), some of which may participate in the chronic regulation of this gene. The glucocorticoid responsiveness of the Nhe3 gene was assessed by fusing its 5' regulatory region to the firefly luciferase reporter gene and then by measuring the expression of the chimeric gene in transiently transfected renal epithelial OK and LLC-PK1 cells. Glucocorticoid treatment significantly increased the luciferase activity of the chimeric gene in both cell lines, thereby indicating that glucocorticoid regulation of Nhe3 is mediated primarily by a transcriptional mechanism.
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PMID:Genomic organization and glucocorticoid transcriptional activation of the rat Na+/H+ exchanger Nhe3 gene. 863 55

We have isolated and sequenced a genomic clone coding for the first three exons and the 5'-flanking region of the human fatty-acid synthase gene. The translation initiation site, ATG, is located in exon II. Primer extension and S1 nuclease analyses showed the presence of three transcription initiation (Ti) sites: Ti I, Ti II, and Ti III. The Ti I site is mapped to the beginning of the untranslated exon I and preceded by a promoter with recognizable TATA and CAAT boxes. The Ti II and Ti III sites are located in intron I, at 60 and 49 nucleotides upstream of the translation initiation site ATG in exon II, respectively. These two Ti sites are preceded by four putative Sp1 boxes, but lack TATA and CAAT boxes. Analysis of luciferase reporter gene expression in transient transfection assays confirmed the existence of two promoters. A 200-base pair 5'-flanking region, which has strong promoter activity comparable with that of the CMV promoter, is considered human fatty-acid synthase promoter I. In a wild-type human fatty-acid synthase-luciferase construct, in which promoter I and intron I are present in their natural configuration, the reporter gene activity is only 1% of that of promoter I. Deletion analysis showed the existence of promoter II, which is located in intron I immediately upstream of the Ti II site. The strength of promoter II is approximately th of that of promoter I in transient transfection assays. Further analysis of reporter gene constructs showed that promoter II inhibited the reporter gene activity of the wild-type construct that contained promoter I and intron I and that the spatial separation of the two promoters is important for this inhibition. A model is proposed based on the possibility that the assembly of transcription complexes on promoter II creates a "roadblock" and reduces the overall expression of the fatty-acid synthase gene by interfering with the progression of transcription from promoter I.
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PMID:Human fatty-acid synthase gene. Evidence for the presence of two promoters and their functional interaction. 866 58

In this study the gene for the murine interleukin-11 receptor alpha chain (IL-11Ralpha) has been characterized. The gene spans 9 kilobase pairs of DNA, and the organization of its 14 exons conforms to the pattern observed for other members of the hematopoietin receptor family. Analysis of the 5' end of the cDNA using 5' RACE showed that the first two exons, designated exons 1a and 1b, are spliced to form alternate transcripts. Transcripts initiating from exon 1b were not found in adult tissues but were present in embryonic stem cells. S1 nuclease and 5' rapid amplification of cDNA ends assays demonstrated multiple major and minor sites of transcription initiation for each exon. The putative promoter regions of both exons lacked TATA boxes, although potential recognition sites for several transcription factors including Sp1, AP1, and AP2 were present. A comparison of the murine and human IL-11Ralpha revealed that the 5' sequence upstream of the major site of transcription initiation site for exon 1b is highly conserved. Northern analysis showed that IL-11Ralpha is expressed in many adult murine tissues. A second IL-11Ralpha-like locus containing a sequence homologous to exons 2-13 was also identified.
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PMID:Structural analysis of the gene encoding the murine interleukin-11 receptor alpha-chain and a related locus. 866 2

cDNA and genomic clones encoding mouse Galbeta1, 3GalNAc-specific GalNAc alpha2,6-sialyltransferase (ST6GalNAc II) were isolated, and the structure organization of the gene was determined. The predicted amino acid sequence is 57.4% identical to the chick ST6GalNAc II sequence but 33.8% identical to the chick ST6GalNA I (GalNAc alpha2, 6-sialyltransferase) sequence. The ST6GalNAc II gene is constitutively expressed in various mouse tissues but highly expressed in lactating mammary gland and adult testis. The gene contains nine exons spanning about 25 kilobases of genomic DNA and encodes a messenger RNA of 1995 nucleotides. Primer extension and S1 nuclease protection analysis of submaxillary gland mRNA showed that the transcription of the ST6GalNAc II gene starts from 68 nucleotides upstream from the translation start site. Characterization of 5'-flanking genomic regions indicated that the Galbeta1,3GalNAc-specific GalNAc alpha2,6-sialyltransferase promoter is embedded in a G+C-rich domain and contains no TATA or CAAT box but has putative binding sites for transcription factors Sp1 and AP-2. Transient transfection experiments involving luciferase reporter genes demonstrated promoter activity in NIH3T3 cells.
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PMID:Molecular cloning and genomic analysis of mouse Galbeta1, 3GalNAc-specific GalNAc alpha2,6-sialyltransferase. 866 27

Alkylations at base nitrogens in DNA are removed by excision repair, the first step of which is catalyzed by the repair enzyme N-methylpurine-DNA glycosylase (MPG). To study regulation of MPG expression, we have cloned the rat MPG promoter. A cosmid clone containing the rat MPG gene was isolated from a library using rat MPG cDNA as a probe. The 5' part of the MPG gene and the nontranscribed 5'-flanking region were isolated and characterized. Transcription start sites of the rat MPG gene were identified by primer extension and S1 nuclease protection analysis of RNA from primary rat hepatocytes. Promoter activity of the 5'-flanking noncoding region was shown by transfection in H4IIE rat hepatoma cells of various genomic MPG fragments cloned in front of the reporter gene chloramphenicol acetyltransferase. The rat MPG promoter does not contain a TATA box, but has a CCAAT sequence element and putative binding sites for the transcription factors Sp1, AP-2, AP-3, Ets-1, PEA3, NF-1, p53, c-Myc, NF-kappa B, and the glucocorticoid receptor. The activity of the rat MPG promoter was found to be inducible by the tumor promoter TPA and UV light, but not to a significant extent by methylating agents and ionizing radiation.
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PMID:Isolation and analysis of inducibility of the rat N-methylpurine-DNA glycosylase promoter. 875 39

The human ATP1AL1 gene belongs to the family of Na,K-ATPase and H,K-ATPase (X,K-ATPases) genes. It encodes a catalytic subunit of hitherto unknown human ouabain-sensitive H,K-ATPase that represents a novel third group of X,K-ATPases distinct from the known Na,K-ATPase and gastric H,K-ATPase. Cloning of the ATP1AL1 gene is described in this report. The exon-intron structure of ATP1AL1 was found to be very similar to that of related genes. It contains 23 exons and spans approximately 32 kb of genomic DNA. All ATP1AL1 exons and 12 of its 22 introns were entirely sequenced. A total of nine Alu repeats were identified in introns. The transcription initiation site was mapped 187 bp upstream of the ATG initiation codon by primer extension and S1 nuclease protection analyses of RNA from human skin and colon. Sequence analysis of the 5'-flanking region (1.48 kb) revealed numerous potential binding sites for transcription factors Sp1 and AP2 and one putative NF-kappa B binding site. The 0.85-kb region from position -484 (5'-flanking region) to position +369 (intron 1) meets the structural criteria of a CpG island. It is suggested that the ATP1AL1 gene contains two poly(A) addition sites that may function in a tissue-specific manner.
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PMID:Genomic organization of the human ATP1AL1 gene encoding a ouabain-sensitive H,K-ATPase. 883 94


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