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 promoter for the gene encoding the low affinity L-arabinose uptake protein in Escherichia coli was studied. The promoter was cloned, sequenced, its transcription start site determined by S1 nuclease mapping, the proteins required for in vitro transcription were determined, and the regulatory protein binding sites located by DNase footprinting. The araE promoter shows no evidence of an operator site upstream from the CRP binding site, but otherwise it is similar to the araBAD promoter.
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PMID:The araE low affinity L-arabinose transport promoter. Cloning, sequence, transcription start site and DNA binding sites of regulatory proteins. 631 8

Sea urchin (Hemicentrotus pulcherrimus) arylsulfatase(Ars) gene contains a long (622 bp) polypyrimidine:polypurine (Pyr-Pur) sequence in its 5' flanking region. The Pyr-Pur sequence inserted into a plasmid was sensitive to S1 nuclease at a low acidic pH (pH 5) when the plasmid was negatively supercoiled. From the distribution pattern of S1 sites in the Pyr-Pur region it is concluded that a (CT)11:(GA)11 tract in this region could adopt an unusual DNA configuration distinct from the usual B-form. Another feature of the Pyr-Pur sequence is that this (CT)11:(GA)11 tract is sandwiched by two oligo(dC):oligo(dG) stretches (G-strings) that are located at almost an equal distance from both ends of the (CT)11:(GA)11 tract. Mobility shift assay and DNase-I footprinting revealed that the gastrula nuclei contain nuclear proteins that interact with two distinct oligo(dG):oligo(dG) tracts (G-strings) in the Pyr-Pur region. The possibility is suggested that G-strings may be related to formation and stabilization of an unusual DNA configuration of a (CT)11:(GA)11 tract.
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PMID:A long polypyrimidine:polypurine sequence in 5' flanking region of arylsulfatase gene of sea urchin embryo. 798 Oct 42

The sequence of the S-adenosyl-L-methionine:trans-caffeoyl-CoA O-methyltransferase (CCoAOMT, EC2.1.1.104) gene, including the 5'-flanking region of 5 kb, was determined from parsley (Petroselinum crispum) plants. The enzyme appears to be encoded by one or two genes, and the ORF is arranged in five exons spaced by introns from 107 to 263 bp in length. The genomic sequence matches the ORF of the cDNA previously reported from elicited parsley cell cultures, showing only three base changes that do not affect the enzyme polypeptide sequence. S1 nuclease protection assays and primer extension analyses with genomic and cDNA templates revealed the transcription start site 67 bp upstream of the translation start codon, indicating a shorter 5'-UTR than reported previously for the transcript. Promoter regulatory consensus elements such as two 'CAAT' boxes and one 'TATA' box were identified at -196, -127 and -31, respectively, relative to the transcription start site, and an SV 40-like enhancer element is located 347 bp upstream. Most notably, three putative cis-regulatory elements were recognized by sequence alignments, which represent motifs recurring in the promoters of several genes of the stress-inducible phenylpropanoid pathway (boxes P, A and L). Transient expression assays with a set of 5'-truncated promoter-GUS fusions show that significant promoter activity is retained in a 354 bp promoter fragment. In vitro DNase 1 footprint experiments and electrophoretic mobilty shift assays (EMSA) identified in this fragment a unique sequence motif with elicitor-inducible trans-factor binding activity, which was unrelated to boxes P, A, or L. This novel cis-regulatory element, designated box E, appears to be conserved in the TATA-proximal regions of other stress-inducible phenylpropanoid genes, and in vitro binding of nuclear protein was confirmed in EMSA assays for such an element from the PAL-1 promoter (-54 to -45). Moreover, the deletion of box E reduced the activity and erased the elicitor-responsiveness of the CCoAOMT promoter in transient expression assays. The results corroborate the proposed physiological function of CCoAOMT in elicited plant cells and may shed new light on the sequential action of trans-active factors in the regulation of phenylpropanoid genes.
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PMID:Structure of the parsley caffeoyl-CoA O-methyltransferase gene, harbouring a novel elicitor responsive cis-acting element. 903 50

We describe a new technique that allows specific visualization of RNA at the electron microscopic level by means of terbium citrate. Under the conditions presented here, terbium binds selectively to RNA and stains nucleoli, interchromatin granules, peri-chromatin fibrils, perichromatin granules, and coiled bodies in the cell nucleus, whereas ribosomes are the only contrasted structures in the cytoplasm. All the cell components contrasted by terbium are known to contain RNA. When ultrathin sections are pretreated with RNase A or nuclease S1 (specific for single-stranded nucleic acids), staining does not occur. Neither DNase nor pronase influences the reaction. We conclude that terbium staining is selective for RNA and especially for single-stranded RNA. The staining can be performed on thin sections of material embedded both in epoxy and in acrylic resins. The technique is not influenced by the aldehyde fixative used and can also be utilized after immunolabeling. The endproduct is very fine and, although weak in contrast, is suitable for high-resolution observations.
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PMID:Fine structural specific visualization of RNA on ultrathin sections. 948 21

Glucose-6-phosphate dehydrogenase (G6PD) activity differs among tissues and, in liver, with the dietary state of the mouse. Tissue-specific differences in G6PD activity in adipose tissue, liver, kidney, and heart were associated with similar differences in the amount of G6PD mRNA. Regulation of mRNA amount by dietary fat was only observed in liver. In mice fed a low-fat diet, the relative amounts of G6PD mRNA were 3:1:1:0.38, respectively, in the four tissues. Further, the amount of precursor mRNA for G6PD in liver, kidney, and heart reflected the amount of mature mRNA in these tissues, suggesting differing transcriptional activity. Our S1 nuclease and primer-extension analyses indicated that the same transcriptional start site is used in liver, kidney, and adipose tissue, resulting in a common 5' end of the mRNA in these tissues. Thus, differential regulation is not attributable to alternate promoter usage. A DNase hypersensitivity analysis of the 5' end of the G6PD gene identified three hypersensitive sites (HS): HS 1 and HS 2 were present in all tissues, whereas HS 3 was liver specific. Thus, regulation of G6PD expression involves both dietary and tissue-specific signals that appear to act via different mechanisms.
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PMID:Structural characterization and tissue-specific expression of the mouse glucose-6-phosphate dehydrogenase gene. 953 8

Nucleic acid sequence-based amplification (NASBA) is a technique that has been previously shown to selectively mediate the detection of RNA in microbial cells. In a series of tests, nucleic acids were extracted from Salmonella enterica serotype Typhimurium and Mycobacterium avium subsp. paratuberculosis, and subjected to four enzymatic treatments prior to NASBA. These enzymatic treatments were DNase, RNase, S1 nuclease, and RNase/S1 nuclease. The results obtained were different for the two bacteria. With S. enterica serotype Typhimurium, RNase and RNase/S1 nuclease abolished the NASBA signal, as expected. But with M. avium subsp. paratuberculosis RNase, S1 nuclease, and RNase/S1 nuclease had no effect on the NASBA signal, whereas DNase treatment abolished it. This indicates that in the latter bacterium, NASBA can detect DNA, and demonstrates the necessity of verifying the nucleic acid origin of a NASBA signal if detection of RNA is objective.
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PMID:Unexpected detection of DNA by nucleic acid sequence-based amplification technique. 1527 85

DNA sequences encoding hypothetical proteins homologous to S1 nuclease from Aspergillus oryzae are found in many organisms including fungi, plants, pathogenic bacteria, and eukaryotic parasites. One of these is the M1 nuclease of Mesorhizobium loti which we demonstrate herein to be an enzymatically active, soluble, and stable S1 homolog that lacks the extensive mannosyl-glycosylation found in eukaryotic S1 nuclease homologs. We have expressed the cloned M1 protein in M. loti and purified recombinant native M1 to near homogeneity and have also isolated a homogeneous M1 carboxy-terminal hexahistidine tag fusion protein. Mass spectrometry and N-terminal Edman degradation sequencing confirmed the protein identity. The enzymatic properties of the purified M1 nuclease are similar to those of S1. At acidic pH M1 is 25 times more active on single-stranded DNA than on double-stranded DNA and 3 times more active on single-stranded DNA than on single-stranded RNA. At neutral pH the RNase activity of M1 exceeds the DNase activity. M1 nicks supercoiled RF-I plasmid DNA and rapidly cuts the phosphodiester bond across from the nick in the resultant relaxed RF-II plasmid DNA. Therefore, M1 represents an active bacterial S1 homolog in spite of great sequence divergence. The biochemical characterization of M1 nuclease supports our sequence alignment that reveals the minimal 21 amino acid residues that are necessarily conserved for the structure and functions of this enzyme family. The ability of M1 to degrade RNA at neutral pH implies previously unappreciated roles of these nucleases in biological systems.
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PMID:Characterization of a periplasmic S1-like nuclease coded by the Mesorhizobium loti symbiosis island. 1652 13

A new virus species of the genus Phytoreovirus was isolated from glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar (Hemiptera: Cicadellidae), in California and designated here as Homalodisca vitripennis reovirus (HoVRV). Extraction of nucleic acid from GWSS adults collected from three Californian populations revealed an array of double-stranded (ds) RNA species that was soluble in 2 M LiCl and resistant to degradation upon exposure to S1 nuclease and DNase. Analysis of nucleic acid samples from single GWSS adults indicated that HoVRV dsRNA accumulated to high titer in individual insects. Double-shelled isometric virus particles purified from GWSS adults resembled those observed in thin sections of GWSS salivary glands by transmission electron microscopy. Purified HoVRV virions contained 12 dsRNA segments that, based on complete nucleotide sequences, ranged in size from 4475 to 1040 bp. Sequence comparisons indicated that the HoVRV dsRNA segments were most closely related (58.5 to 43.7% nt sequence identity) to the corresponding genome segments of Rice dwarf virus (RDV). Each HoVRV dsRNA segment encoded a single open reading frame (>300 nts) except for segment 11, which appears to be dicistronic. Terminal nucleotide sequences of HoVRV positive-sense RNAs were similar to other phytoreoviruses (GGCG or GGCA at the 5'-end and UGAU or CGAU at the 3'-end) with adjacent imperfect inverted repeats potentially able to base pair. Phylogenetic analyses of the RNA-directed RNA polymerase (encoded by segment 1) and the outer capsid protein (encoded by segment 8) confirmed placement of HoVRV as a species of the genus Phytoreovirus sharing a most recent common ancestor with RDV. Reverse transcription-polymerase chain reaction assays revealed that HoVRV infection of GWSS in California was common and that the virus also occurred in GWSS populations from the Carolinas and Texas.
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PMID:A new Phytoreovirus infecting the glassy-winged sharpshooter (Homalodisca vitripennis). 1923 72

Pseudocercospora griseola (Sacc.) Crous & Braun is a widespread fungal phytopathogen that is responsible for angular leaf spot in the common bean (Phaseolus vulgaris L.). A number of fungal phytopathogens have been shown to harbour mycoviruses, and this possibility was investigated in populations of Pseudocercospora griseola. The total nucleic acid extracts of 61 fungal isolates were subjected to agarose gel electrophoresis. Small fragments (800-4800 bp) could be identified in 42 of the samples. The presence of dsRNA in isolate Ig838 was confirmed by treatment of total nucleic acid with DNase, RNase A, and nuclease S1. Transmission electron microscopy revealed the presence of viral-like particles 40 nm in diameter in the mycelia of 2 fungal isolates, namely 29-3 and Ig838. The transmission of dsRNA by means of conidia was 100% for isolate 29-3, but there was loss of 1-6 fragments of dsRNA in monosporic colonies of isolate Ig848. Cycloheximide treatment failed to inhibit the mycovirus in isolate 29-3, but proved efficient in the elimination of the 2.2, 2.0, 1.8, 1.2 and 1.0 kb fragments in 2 colonies of isolate Ig848. The occurrence of a mycovirus in Pseudocercospora griseola was demonstrated for the first time in the present study.
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PMID:Mycovirus in Pseudocercospora griseola, the causal agent of angular leaf spot in common bean. 2055 97


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