Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

Fatty acid synthase is regulated by diet and hormones, with regulation being primarily transcriptional. In chick embryo hepatocytes in culture, triiodothyronine stimulates accumulation of enzyme and transcription of the gene. Since the 5'-flanking region of this gene is likely involved in hormonal regulation of its expression, we have isolated and partially characterized an avian fatty acid synthase gene. A genomic DNA library was constructed in a cosmid vector and screened with cDNA clones that contained sequence complementary to the 3' end of goose fatty acid synthase mRNA. A genomic clone (approximately 35 kilobase pairs (kb] was isolated, and a 6.5-kb EcoRI fragment thereof contained DNA complementary to the 3' noncoding region of fatty acid synthase mRNA. Additional cosmid libraries were screened with 5' fragments of previously isolated genomic clones, resulting in the isolation of five overlapping cosmid DNAs. The entire region of cloned DNA spans approximately 105 kb. Exon-containing fragments were identified by hybridization with end-labeled poly(A)+ RNA and by hybridization of labeled exon-containing genomic DNA fragments to fatty acid synthase mRNA. A new set of cDNA clones spanning approximately 3.2 kb was isolated from a lambda-ZAP goose liver cDNA library using the 5'-most exon-containing fragment of the 5'-most genomic DNA clone. This region of mRNA contains a 5'-untranslated sequence and a continuous open reading frame which includes a region that codes for the essential cysteine of the beta-ketoacyl synthase domain. The entire fatty acid synthase gene spans about 50 kb. The 5' 15 kb of the gene contain 7 exons. S1 nuclease and primer extension analyses were used to identify a single site for initiation of transcription, 174 nucleotides upstream from the putative translation initiation codon. Putative "TATA" and "CCAAT" boxes are located 28 and 60 base pairs (bp), respectively, upstream of the site of initiation of transcription. The 5'-flanking 597 bp of DNA contains G/C-rich sequences including several "GC" boxes corresponding to binding sites for the nuclear transcription factor Sp1. Putative sites for AP-2, C/EBP, and the triiodothyronine and glucocorticoid receptors also were found in this region. A chimeric DNA, containing approximately 1.6 kb of 5'-flanking sequence and 139 bp of untranslated sequence of the goose fatty acid synthase gene ligated to the bacterial chloramphenicol acetyl-transferase (CAT) gene, was transfected into chick embryo hepatocytes in culture. Cells treated with triiodothyronine contained increased chloramphenicol acetyltransferase and fatty acid synthase activities.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Isolation and partial characterization of the gene for goose fatty acid synthase. 170 26

A defect in the E1 beta subunit of the branched chain alpha-keto acid dehydrogenase (BCKDH) complex is one cause of maple syrup urine disease (MSUD). In an attempt to elucidate the molecular basis of MSUD, we isolated and characterized the cDNA of the E1 beta subunit of BCKDH. Using the cDNA as a probe, a chromosomal gene related to E1 beta subunit of human BCKDH was isolated from human gene libraries. The gene of E1 beta subunit is over 100 kilobases long and is split into 10 exons. All of the splice donor and acceptor sites conform to the GT/AG rule. The transcription initiation site was determined by nuclease S1 mapping and primer extension and was located 47 bases upstream from the initiation codon. A "CAAT" box and its reverse complement sequences were present at 39 bases and 75 bases upstream from the cap site, but there was no "TATA" box-like sequence. There were three sets of sequences resembling the transcription factor Sp1-binding sites and two sets of sequences resembling the enhancer core sequence. We also analyzed the chromosomal localization of the gene for the E1 beta subunit of BCKDH. The gene was mapped to chromosome 6. Knowledge of the gene structure of human BCKDH E1 beta subunit will facilitate further studies on the expression and regulation of this gene and provide necessary information for analyses of mutations in patients with MSUD.
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PMID:Structural organization and chromosomal localization of the gene for the E1 beta subunit of human branched chain alpha-keto acid dehydrogenase. 186 Aug 67

We have determined the sequence of the 5'-flanking region and first three exons of the human Na,K-ATPase alpha 1 gene, ATP1A1. Primer extension and S1 nuclease protection analyses of RNA from human kidney, brain, and skeletal muscle indicate that transcription initiates 273 nucleotides upstream of the translation start site. The promoter region contains a potential TATA box at position -27 relative to the transcription initiation site; however, no CCAAT sequence is observed. The 5'-untranslated and 5'-flanking regions are G + C rich. Five sequence elements exhibiting similarity to binding sites for the transcription factor Sp1 are located within the 5'-flanking region. This region also contains potential binding sites for the transcription factors AP-1, AP-2, AP-3, and NF-1, as well as a site which exhibits perfect identity to an 8-bp sequence element important for calcium induction. A comparison of the 5'-flanking region of the alpha 1 and alpha 2 genes reveals differences in potential transcription factor and hormone receptor binding sites which may be important in mediating the tissue- and developmental stage-specific expression of these genes. We have also identified an intragenic DNA probe which detects a restriction fragment length polymorphism at the alpha 1 locus. This marker should facilitate genetic linkage studies designed to evaluate the role of the sodium pump in human disease.
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PMID:The human Na, K-ATPase alpha 1 gene: characterization of the 5'-flanking region and identification of a restriction fragment length polymorphism. 197 Mar 26

Prolidase (peptidase D) catalyzes hydrolysis of the di- and tripeptide with carboxyl-terminal proline and plays an important role in recycling proline in various cells and tissues. By using human prolidase cDNA as a probe, a chromosomal gene related to prolidase was isolated from human gene libraries. The human prolidase gene is over 130 kilobases long and is split into 15 exons. All of the splice donor and acceptor sites conform to the GT/AG rule. The transcription initiation site was determined by nuclease S1 mapping and primer extension and was located 131 bases upstream from the initiation codon. A "CAAT" box-like sequence was present 67 bases upstream from the cap site, but there was no "TATA" box-like sequence. There were seven sets of sequences resembling the transcription factor Sp1 binding sites. Four were upstream from the cap site, and three were downstream. We also analyzed findings in patients with prolidase deficiency with respect to major gene re-arrangement. Several hundred base deletions, including the 14th exon, were identified. Knowledge of the gene structure of human prolidase will facilitate further studies on the expression and regulation of this gene and provide necessary information for analyses of mutations in patients with this deficiency.
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PMID:Structural organization of the gene for human prolidase (peptidase D) and demonstration of a partial gene deletion in a patient with prolidase deficiency. 197 7

The carbamoylphosphate synthetase-aspartate transcarbamylase-dihydroorotase (CAD) gene encodes a tri-functional protein catalyzing the first three steps in de novo pyrimidine biosynthesis. Studies correlating CAD gene expression with cellular proliferation indicate the importance of understanding the regulation of the CAD gene. As a first step, the structure of the promoter region of the Syrian hamster CAD gene has been determined. Sequence analysis of 1671 base pairs of DNA revealed that the CAD promoter region is very GC rich. Primer extension analysis indicated that the transcription initiation site of the CAD gene is downstream from two GC boxes (consensus binding sites for the transcription factor Sp1). There is no TATA box appropriately spaced upstream from the transcription initiation site. Using RNase protection mapping, S1 nuclease analysis, and comparison to consensus splice donor/acceptor sites, the 5' end of the CAD gene has been determined to consist of a 241-base pair first exon, a 187-base pair first intron, a 140-base pair second exon, and a second intron that extends at least three kilobase pairs. Using conditions optimized for this GC-rich promoter, accurate transcription can be achieved in vitro. Analysis of CAD promoter deletions indicated that sequences extending only 114 base pairs upstream and 225 base pairs downstream from the transcription initiation site are sufficient for accurate and efficient transcription in vitro. DNase I footprinting reactions using this promoter fragment have identified three regions that bind proteins in a HeLa nuclear extract.
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PMID:Characterization of the 5' end of the growth-regulated Syrian hamster CAD gene. 198 61

The structural organization of the entire nuclear gene (NMDMC) encoding the mitochondrial (mt) NAD-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase enzyme (NMDMC) was determined by analysis of clones obtained from a lambda EMBL3 murine genomic DNA library. The gene is approx. 13 kb in length and contains eight exons and seven introns. All exon/intron splice junctions follow the GT/AG rule. The amino acid presequence, which is essential for transport of the NMDMC enzyme precursor into mt, is encoded almost entirely in the first exon. Two major transcriptional start points (tsp), located 33 and 75 nucleotides upstream from the AUG start codon, were revealed by S1 nuclease mapping and RNase protection analyses. The immediate 5'-flanking region of the first exon contains one CAAT box, a TATA-like box and three sites homologous to the consensus sequence for the binding of transcription factor Sp1.
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PMID:Structural organization of the murine gene encoding NAD-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase. 199 93

Genomic clones containing the 5'-terminal portion of the human CRE-BP1 gene that encodes transcriptional regulator binding to the cyclic AMP response element (CRE) were isolated. Multiple transcriptional start sites in the promoter region were identified by nuclease S1 mapping and primer extension analysis. By DNase I footprinting with use of purified transcription factor Sp1 and nuclear extracts prepared from HeLa cells, 11 Sp1-binding sites, two CCAAT sequences, two CREs, and three unknown factor recognition elements were found. Transfection of chimeric chloramphenicol acetyltransferase plasmids containing various deletions of the promoter into CV-1 cells indicated that the region between nucleotides -50 and 90, which contained three Sp1-binding sites and one CRE, was sufficient for basal promoter activity. These results suggest that multiple sequence-specific DNA-binding proteins may control the expression of the CRE-BP1 gene, although Sp1 seems to be important for the basal promoter activity.
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PMID:Promoter region of the human CRE-BP1 gene encoding the transcriptional regulator binding to the cyclic AMP response element. 214 72

The structural organization of the X-linked gene for the E1 alpha subunit of the human pyruvate dehydrogenase complex has been determined by restriction endonuclease mapping and DNA sequence analysis of overlapping genomic clones. The gene is approximately 17 kilobase pairs long. It contains 11 exons ranging from 61 to 174 base pairs and introns ranging from 600 base pairs to 5.7 kilobase pairs. All the splice donor and acceptor sites conform to the GT/AG rule. The transcription initiation site was determined by S1 nuclease mapping. The DNA sequence around this site is very GC-rich. A "TATA box"-like sequence and a "CAAT box"-like sequence are present 24 and 113 bases upstream from the cap site, respectively. Also upstream from the cap site are several sets of inverted repeats, direct repeats, several sequences resembling the transcription factor Sp1 binding site, a glucocorticoid-responsive element, and two cAMP receptor binding sites.
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PMID:Structural organization of the gene for the E1 alpha subunit of the human pyruvate dehydrogenase complex. 274 44

The murine urokinase-type plasminogen activator (uPA) gene has been isolated from a BALB/c liver DNA cosmid library and its nucleotide sequence established. The gene is organized into 11 exons comprising 34.7% of the 6710 base pair (bp) region spanning the interval between the presumed transcription initiation and polyadenylation sites. The transcription initiation site is flanked by common RNA polymerase II promoter elements, including a TATA box and a potential transcription factor Sp1 binding site. A large polypurine tract of the structure (AG)22(AGGG)16(AG)28 is located 79 bp upstream of the 5'-terminus. It was highly sensitive to the single-strand-specific nuclease S1, suggesting a non-B-DNA conformation of unknown significance. Consistent with the well-documented influence of adenosine cyclic 3',5'-phosphate (cAMP) on uPA gene expression, there is a dodecanucleotide homologous to proposed regulatory sequences identified in other cAMP-modulated genes. Comparison of the murine uPA gene to the previously described porcine and human uPA genes revealed an unusually high degree of evolutionary (interspecies) sequence conservation that was not limited to exons but included introns and flanking sequences as well.
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PMID:The murine urokinase-type plasminogen activator gene. 283 40

The epidermal growth factor (EGF) receptor is the functional target of the mitogen EGF and the cellular homolog of the avian erythroblastosis virus erbB oncogene product. Regulation of expression of the proto-oncogene encoding the EGF receptor can be elucidated by studying the structure and function of the gene promoter outside the confines of the cell. Previously, we reported the isolation of the human EGF receptor gene promoter. The promoter is highly GC rich, contains no TATA or CAAT box, and has multiple transcription start sites. An S1 nuclease-sensitive site has now been found 80 to 110 base pairs (bp) upstream from the major in vivo transcription initiation site. Two sets of direct repeat sequences were found in this area; both conform to the motif TCCTCCTCC. When deletion mutations were made in this region of the promoter by using either Bal 31 exonuclease or S1 nuclease, we found that in vivo activity dropped three- to fivefold, on the basis of transient-transfection analysis. Examination of nuclear protein binding to normal and mutated promoter DNAs by gel retardation analysis and DNase I footprinting revealed that two specific factors bind to the direct repeat region but cannot bind to the S1 nuclease-mutated promoter. One of the specific factors is the transcription factor Sp1. The results suggest that these nuclear trans-acting factors interact with the S1 nuclease-sensitive region of the EGF receptor gene promoter and either directly or indirectly stimulate transcription.
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PMID:Modulation of epidermal growth factor receptor proto-oncogene transcription by a promoter site sensitive to S1 nuclease. 284 30


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