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)

The levels of chlorophyll a/b-binding protein (Cab) gene polysomal poly(A)+ mRNA were quantitated throughout the development of Glycine max L. Cab mRNAs were abundant in young expanding leaves, representing 6.1% of the leaf mRNA population. Lower Cab mRNA levels were present in embryos, stems, and cotyledons of developing seedlings; the lowest levels were found in roots where they accounted for 0.04% of the polysomal poly(A)+ mRNA of this organ. To determine the contribution of different members of the Cab gene family to the Cab mRNA populations, a quantitative S1 nuclease reconstruction assay was developed. Cab3, Cab4, and Cab5 mRNAs were detected in all stages examined during soybean development but their levels underwent differential changes. Cab3 encodes the most abundant Cab mRNA in young leaves, developing embryos, and in Stage VII cotyledons from the developing soybean seedling. The levels of Cab mRNAs were compared to the levels of ribulose-1,5-bisphosphate carboxylase small subunit gene mRNA and differences in their patterns of accumulation were noted. Collectively these data indicate that during soybean embryogenesis developmental control mechanisms supersede light-regulatory signals.
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PMID:Chlorophyll a/b-binding protein genes are differentially expressed during soybean development. 135 68

The bean rust fungus, Uromyces appendiculatus, undergoes thigmotropic differentiation to produce infection structures. Six differentiation-specific genes have been isolated and one, INF24, has been characterized [Bhairi et al., Gene 81 (1989) 237-243]. Here, we report the structure of a second gene, INF56, which was subcloned on a 2.6-kb fragment and sequenced. The location of the 1.0-kb INF56 transcript was determined by S1 nuclease protection and primer extension. A TATA box was found 38 bp upstream and a CAAT box 130 bp upstream from the major transcription start point (tsp). The gene contains two open reading frames: ORF2 is nested within ORF1; they share a 67-bp intron. ORF1 encodes a 14.1-kDa polypeptide which has an amino acid sequence rich in Gly, Pro and Ser. It has sequence similarity to a functional domain (V2) of mammalian cytokeratin type II. ORF2 encodes a 10.1-kDa polypeptide which is rich in Pro. It shares similarity with the cell-surface recognition region of chicken fibronectin. Hybrid selection and in vitro translation of the INF56 mRNA yielded two polypeptides of 15.5 and 23 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. INF56 is constitutively expressed at a low level, but the abundance of its steady-state transcript is upshifted 4.5 h after spore hydration during the period that infection structures are formed.
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PMID:Characterization of INF56, a gene expressed during infection structure development of Uromyces appendiculatus. 154 77

By screening of an Escherichia coli plasmidic library using antibodies against aspartyl-tRNA synthetase (AspRS) several clones were obtained containing aspS, the gene coding for AspRS. We report here the nucleotide sequence of aspS and the corresponding primary structure of the aspartyl-tRNA synthetase, a protein of 590 amino acid residues with a Mr 65,913, a value in close agreement with that observed for the purified protein. Primer extension analysis of the aspS mRNA using reverse transcriptase located its 5'-end at 94 nucleotides upstream of the translation initiation AUG; nuclease S1 analysis located the 3'-end at 126 nucleotides downstream of the stop codon UGA. Comparison of the DNA-derived protein sequence with known aminoacyl-tRNA sequences revealed important homologies with asparaginyl- and lysyl-tRNA synthetases from E.coli; more than 25% of their amino acid residues are identical, the homologies being distributed preferencially in the first part and the carboxy-terminal end of the molecule. Mutagenesis directed towards a consensus tetrapeptide (Gly-Leu-Asp-Arg) and the carboxy-terminal end showed that both domains could be implicated in catalysis as well as in ATP binding.
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PMID:Aspartyl-tRNA synthetase from Escherichia coli: cloning and characterisation of the gene, homologies of its translated amino acid sequence with asparaginyl- and lysyl-tRNA synthetases. 212 59

A 2.4-kb genomic clone, which encodes the precursor of a snake neurotoxin, erabutoxin c, was isolated from the liver of the sea snake, Laticauda semifasciata. The erabutoxin c gene is about 1.2 kb long and consists of three exons and two introns. The first intron is found at the position corresponding to the signal peptide between amino acid residues 18(Leu) and 19(Gly). The second intron is located at the position corresponding to the central loop of the mature toxin, between amino acid residues 33(Arg) and 34(Gly). A TATA box consensus sequence, characteristic of promoter regions, is identified 29-33 nucleotides upstream from the transcription-initiation site identified by nuclease S1 analysis. The erabutoxin c cDNA nucleotide sequence, deduced from the present work, is compared with the cDNA sequences of erabutoxins a and b reported previously. Replacements are found at three positions, two of which correspond to the amino acid replacements found among the toxins, while the other is silent.
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PMID:Structure of the snake short-chain neurotoxin, erabutoxin c, precursor gene. 224 84

Murine macrophage cell lines and resident macrophages showed various levels of expression of the murine osteopontin (OP) gene, and macrophage stimulating agents were found to enhance transcription of the gene with kinetics which are unique for each stimulator. The organization of the murine OP gene was determined. The gene comprises six exons and five introns and spans approximately 4.8 kilobases. Exon 1 contains the 16 amino acids of the leader sequence. Exons 2, 3, 4, 5, and 6 encode 12, 27, 14, 94, and 129 amino acid residues, respectively. Exon 5 encodes regions containing 10 consecutive Asp amino acid residues and a Gly-Arg-Gly-Asp-Ser peptide. Exon 6 encodes the C-terminal half of OP and contains no 15- and 54-base pair nucleotide sequences which are deleted in murine OP cDNA compared to that of rat OP cDNA. Since Southern blot analysis indicated that the OP gene is a single copy, it is obvious that the murine OP cDNA has the sequence previously determined (Miyazaki, Y., Setoguchi, M., Yoshida, S., Higuchi, Y., Akizuki, S., and Yamamoto, S. (1989) Nucleic Acids Res. 17, 3298). A comparison with the cDNA sequences reported previously suggested the presence of nucleotide sequence polymorphisms. The 5' end of the murine OP gene was defined by primer extension and S1 nuclease mapping. Sequence analysis of the 5'-flanking DNA revealed the presence of many potential regulatory motifs.
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PMID:The mouse osteopontin gene. Expression in monocytic lineages and complete nucleotide sequence. 238 63

The +1 site for transcription initiation of the inducible 23 S rRNA adenine methylase encoded by plasmid pE194 was determined experimentally by nuclease S1 mapping of mRNA synthesized in vivo, and by nuclease T1 mapping of (5'-gamma-32P)-end-labeled transcripts synthesized in vitro. By partial digestion of the in vitro transcripts using S1 and cobra venom nuclease as probes of mRNA conformation, the analysis was extended to reveal single-stranded and double-stranded regions, respectively, which correspond to the critical stems and loops postulated for active and inactive conformations of the nascent mRNA. According to the model for induction, the transition from inactive to active conformation involves disruption of mRNA secondary structure which, in turn, is predicated on protracted occupancy by ribosomes complexed with erythromycin of one of the critical stem sequences. Ribosome occupancy of the critical stem sequence is due to the presence of an open reading frame that encodes part of a 19 amino acid residue "leader" peptide. The existence of this peptide, deduced from the nucleotide sequence of the control region upstream from the methylase structural gene, was demonstrated in vivo as part of a translational fusion with Escherichia coli beta-galactosidase in which the first four amino acid residues of the N-terminal sequence of the fusion protein, analyzed directly by the microsequencing method, were found to comprise N-terminal amino acids 2 through 5, Gly-Ile-Phe-Ser, predicted for the leader peptide.
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PMID:Messenger RNA from Staphylococcus aureus that specifies macrolide-lincosamide-streptogramin resistance. Demonstration of its conformations and of the leader peptide it encodes. 241 56

The complete nucleotide sequences of the 1.2-kilobase HindIII fragments which contain the pilin genes of two independently isolated strains of Pseudomonas aeruginosa (PAK and PA103) have been determined and compared to that of strain PA01 (Sastry, P. A., Finlay, B. B., Pasloske, B. L., Paranchych, W., Pearlstone, J. R., and Smollier, L. B. (1985) J. Bacteriol. 164, 571-577). The fragments share extensive regions of homology, including the 5'- and 3'-flanking sequences as well as the 5' end of the pilin gene. The most highly diverged segments of the pilin genes are those which encode the variable carboxyl-terminal region of the pilin polypeptides. The pilin polypeptides each contain a 6-amino acid amino-terminal leader peptide (Met-Lys-Ala-Gln-Lys-Gly) and are nearly identical in the following 60 amino acids. The carboxyl-terminal portion of the pilin polypeptides contain extensive regions of divergence in their amino acid sequences, although hydropathicity analysis of the pilin polypeptides indicated that they are structurally similar. The transcriptional initiation site of the PAK pilin gene has been determined by S1 nuclease mapping. The promoter region at -10 and -35 base pairs from the transcriptional initiation site shows no significant homology to the consensus Escherichia coli promoter, but the -12 and -24 regions show a high degree of homology to promoters which require the ntrA gene product for transcription. Several other Pseudomonas promoters and the promoters of the homologous pilin genes from other bacterial species also share homology to this sequence.
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PMID:Nucleotide sequence and transcriptional initiation site of two Pseudomonas aeruginosa pilin genes. 243 Sep 61

Two overlapping human genomic clones that encode a short-chain collagen, designated alpha 1(XIII), were isolated by using recently described cDNA clones. Characterization of the cosmid clones that span approximately equal to 65,000 base pairs (bp) of the 3' end of the gene established several unusual features of this collagen gene. The last exon encodes solely the 3' untranslated region and it begins with a complete stop codon. The 10 adjacent exons vary in size from 27 to 87 bp and two of them are 54 bp. Therefore, the alpha 1-chain gene of type XIII collagen has some features found in genes for fibrillar collagens but other features that are distinctly different. Previous analysis of overlapping cDNA clones and nuclease S1 mapping of mRNAs indicated one alternative splicing site causing a deletion of 36 bp from the mature mRNA. The present study showed that the 36 bp is contained within the gene as a single exon and also that the gene has a 45-bp -Gly-Xaa-Xaa- repeat coding exon not found in the cDNA clones previously characterized. Nuclease S1 mapping experiments indicated that this 45-bp exon is found in normal human skin fibroblast mRNAs. Accordingly, the data demonstrate that there is alternative splicing of at least two exons of the type alpha 1(XIII)-chain gene.
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PMID:Gene structure for the alpha 1 chain of a human short-chain collagen (type XIII) with alternatively spliced transcripts and translation termination codon at the 5' end of the last exon. 245 7

Gastrin-releasing peptide (GRP), the mammalian homolog of the amphibian peptide bombesin, is encoded in man by a single gene located on chromosome 18. Restriction enzyme and DNA sequence analyses establish that the gene is 10 kilobases in size with two introns of 4.8 and 3.9 kilobases. Exon 1 encodes the 5'-untranslated region, the signal peptide, and the first 23 amino acids of GRP. Exon 2 encodes the remaining three complete amino acids of GRP and the first 74 amino acids of the GRP carboxy-terminal extension peptide. Hence, intron 1 interrupts the coding region of the bioactive portion of GRP between the first and second nucleotides for Gly, the 24th amino acid of GRP. Exon 3 encodes the remainder of the GRP-extension peptide and the 3'-untranslated region. Two GC-rich, potential regulatory sequences and a sequence associated with regulation by cAMP lie between the CAAT and TATA boxes; the primary transcriptional start site is located 30 bases downstream from the TATA box. The second intron has an alternate donor site at its 5'-end and an alternate acceptor site at its 3'-end. S1 nuclease mapping demonstrates that differential RNA splicing using these sites results in the similar expression of three GRP mRNAs in GRP-containing neurons (in stomach and brain) as well as in GRP-containing neuroendocrine cells (fetal lung). In addition, the pattern of RNA splicing is similar between normal tissue and neoplastic tissue (small cell carcinoma of the lung and medullary carcinoma of the thyroid).
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PMID:Analysis of the gene and multiple messenger ribonucleic acids (mRNAs) encoding human gastrin-releasing peptide: alternate RNA splicing occurs in neural and endocrine tissue. 284 May 64

The gene encoding the beta-subunit of rat thyrotropin (TSH beta) has been isolated and characterized. Blot hybridization of restriction enzyme digests of rat genomic DNA suggests that the gene is present in a single copy. The transcriptional unit is 4.9 kilobases in size representing 3 exons interrupted by 2 introns of 3.9 and 0.4 kilobases. Its nucleotide sequence reveals that the locations of the exon/intron junctions are one nucleotide upstream from the translational start and between codons +34 and +35. The location of the second intron is apparently strictly conserved among the glycoprotein hormone beta-subunit genes being four codons downstream from a region encoding a consensus sequence: Cys-Ala-Gly-Tyr. Using S1 nuclease mapping and oligonucleotide-primed reverse transcription of normal and thyroidectomized rat pituitary mRNA, two transcriptional start sites were identified in the rat TSH beta gene that are 28 and 71 nucleotides upstream from the translational start site. The level of TSH beta mRNA containing the downstream site is altered by thyroidal status whereas the other mRNA utilizing the upstream cap site appears to be constitutively expressed. Characteristic promoter elements are present in the 5'-flanking region including TATAAA or Goldberg-Hogness consensus regions which are present 29 and 26 bases upstream from the respective starts of transcription. Also, several CAAT boxlike sequences are located between 95 and 300 bases upstream from the start of translation. Isolation and characterization of the gene encoding the TSH beta gene will facilitate the study of the molecular mechanisms by which hormones regulate TSH beta gene expression.
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PMID:Isolation and characterization of the rat thyrotropin beta-subunit gene. Differential regulation of two transcriptional start sites by thyroid hormone. 302 91


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