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Enzyme
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Query: EC:3.1.27.5 (
RNase
)
17,967
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
T4 Species I RNA, a molecule 140 nucleotides in length with some structural features very much like a tRNA, is specifically cleaved by an enzymatic activity in Escherichia coli extracts to give three segments with 19, 48 and 73 nucleotides. We report the purification and characterization of the E. coli
RNase
which cleaves two 3' phosphodiester bonds of T4 Species I RNA. This reaction has many properties in common with those catalyzed by E. coli RNase III, although the optimal
salt
conditions for T4 Species I RNA cleavage differ significantly from those for other RNase III-catalyzed reactions. The reaction is not catalyzed by extracts from an E. coli strain lacking RNase III activity. Furthermore, T4 Species I RNA is cleaved by highly purified E. coli RNase III to yield the same three specific fragments. We conclude that this specific cleavage is due to the action of RNase III, and that the requirement for lower ionic strength may reveal further important properties about this RNA processing enzyme.
...
PMID:Cleavage of T4 species I ribonucleic acid by Escherichia coli ribonuclease III. 78 26
Bacteriophage T4-coded gene 32-protein is an essential component of the T4 replication and recombination systems. Alberts and co-workers (Alberts, B.M., Amodio, F.J., Jenkins, M., Gutmann, E.D., and Ferris, F.L. (1968) Cold Spring Harbor Symp. Quant. Biol. 33, 289-305) have shown that the major physiological activity of the protein involves preferential and cooperative binding to single-stranded DNA. In this paper, the physiochemical parameters characterizing this "melting" protein system are quantitatively determined. Boundary sedimentation velocity experiments are used to measure the interaction of gene 32-protein with native DNA. The binding is shown to be non-cooperative and involves an overlapping site size (nh) of approximately 10 nucleotide residues (or approximately 5 nucleotide pairs). In analogy with the
ribonuclease
results (Jensen, D.E., and von Hippel, P.H. (1976) J. Biol. Chem. 251, 7198-7214), the logarithm of the association constant (Kh) is found to be linerarly related to log [Na+]. The binding of gene 32-protein to denatured (single-stranded) DNA involves appreciable distortion of the polynucleotide backbone from the unliganded conformation; binding totally unstacks the bases of both ribose- and deoxyribose-containing polynucleotides at 10 degrees, and results in a hyperchromic change exceeding that which can be induced by heating. This hyperchromism induced in poly(dA) on binding gene 32-protein under low
salt
(tight binding) conditions is used to determine a value of nc (the single-stranded DNA site size) of approximately 6.7 nucldotide residues per protein. In addition, gene 32-protein binding to single-stranded polynucleotide induces an unusual circular dichroic spectrum characterized principally by a marked decrease in the magnitude of the positive CD band centered at approximately 265 nm. This spectral change is attributed to significant uncoupling of the transition moments of the vicinal bases of the single-stranded polynucleotide on gene 32-protein binding, in accord with the ultraviolet hyperchromism observed. Binding of gene 32-protein to double helical DNA has virtually no effect on the spectral properties of this conformation...
...
PMID:DNA "melting" proteins. II. Effects of bacteriophage T4 gene 32-protein binding on the conformation and stability of nucleic acid structures. 79 45
Double-stranded RNAs from Penicillium chrysogenum virus have been treated with RNAse III, pancreatic RNAse A and RNAse T1 and the degradation of the RNAs has been studied under different conditions. It was found that only the two former enzymes cut across both strands, RNase T1 cannot cleave double strands. RNase III was shown to digest double-stranded RNA by a two step process: an initial phase of specific cleavage is followed by random degradation. In the first phase the enzyme exhibited a definite preference for some specific base pattern. Partial or complete degradation with
pancreatic RNase
A could also be achieved in media with high
salt
concentration provided that the enzyme: substrate ratio was increased together with the
salt
concentration. By combining different assay techniques, the process of degradation was followed from the early stages to complete digestion and the breakdown products were characterised. It is suggested that a structural change in the enzyme molecules enables them to act on double-stranded RNA. RNAse T1, being unable to cleave double strands, provides a useful tool for studying the secondary structure of RNA molecules. Treatment with different nucleases yielded some new information on the structure of different RNA species in Penicillium stoloniferum virus.
...
PMID:Action of nucleases on double-stranded RNA. 81 98
Cells of Bacillus subtilis heated in high concentrations of sodium dodecyl sulfate (5%) and then washed free of detergent with a hot
salt
solution (80 C) become structurally reorganized into regions of densely compacted cytoplasm (termed zebras) and regions of sparsely filled material (termed spaces). Size distribution studies of zebras indicate that division-suppressed mutants and wild-type cells both yield zebras of comparable length. Similarly the lengths of zebras found in populations emerging from spores are uniform in one-, two-, three-, and four-zebra-containing cells. In contrast, the length of spaces is slightly larger than that of zebras and is unusually large in two-zebra-containing cells. The locations of zebras and spaces along cell length have been studied in spore out-growth populations. A statistical procedure developed previously in genome location investigations was used to analyze the location of zebras along cell length. The data indicate that as cells elongate, new sites arise where the cell contents are strongly bound to the cell surface. Within filament populations produced by division-suppressed mutants there is a linear relationship of mean filament length and zebra number per filament. These data indicate that cytoplasm in filaments with no obvious structural compartmentalizations may be organized into units associated with particular regions of cell surface. The attachment of cell contents to the cell surface may involve deoxyribonucleic acid. Zebra-containing cells digested with proteolytic enzyme and
ribonuclease
are converted to cells that contain a crystalline-like granule fixed at the location of each zebra. Exposure to deoxyribonuclease mobilizes these granules within the cell wall.
...
PMID:Cellular organization of Bacillus subtilis: sodium dodecyl sulfate-induced cell partitioning into zebra structures. 82 Jun 87
Although the bulk of RNA synthesized in vitro by vaccinia virus is 8 to 12 S, a small amount of high molecular weight RNA can be detected. This RNA is virion-associated and is not extruded from the virus as high molecular weight RNA. It is sensitive to
pancreatic RNase
digestion in high
salt
, has a density in neutral CS2SO4 of 1.68 g ml-1 and remains large after digestion with DNase or denaturation in dimethyl sulfoxide. In the presence of high concentrations of virus in the in vitro RNA polymerase reaction, pulse-labeling experiments indicate an RNA sedimenting heterogeneously between 20 and 30 S. Pulse-chase experiments indicate that a fraction of this high molecular weight RNA can be chased into RNA sedimenting at 8 to 12 S. Cleavage into smaller fragments is not dependent on continued RNA synthesis but does require ribonucleoside triphosphates. In the presence of ethidium bromide, the RNA is not cleaved.
...
PMID:In vitro synthesis of a high molecular weight virion-associated RNA by vaccinia. 83 1
The lateral mobility of ribosomes bound to rough endoplasmic reticulum (RER) membranes was demonstrated under experimental conditions. High-
salt
-washed rough microsomes were treated with
pancreatic ribonuclease
(
RNase
) to cleave the mRNA of bound polyribosomes and allow the movement of individual bound ribosomesmfreeze-etch and thin-section electron microscopy demonstrated that, when rough microsomes were treated with
RNase
at 4 degrees C and then maintained at this temperature until fixation, the bound ribosomes retained their homogeneous distribution on the microsomal surface. However, when
RNase
-treated rough microsomes were brought to 24 degrees C, a temperature above the thermotropic phase transition of the microsomal phospholipids, bound ribosomes were no longer distributed homogeneously but, instead, formed large, tightly packed aggregates on the microsomal surface. Bound polyribosomes could also be aggregated by treating rough microsomes with antibodies raised against large ribosomal subunit proteins. In these experiments, extensive cross-linking of ribosomes from adjacent microsomes also occurred, and large ribosome-free membrane areas were produced. Sedimentation analysis in sucrose density gradients demonstrated that the
RNase
treatment did not release bound ribosomes from the membranes; however, the aggregated ribosomes remain capable of peptide bond synthesis and were released by puromycin. It is proposed that the formation of ribosomal aggregates on the microsomal surface results from the lateral displacement of ribosomes along with their attached binding sites, nascent polypeptide chains, and other associated membrane proteins; The inhibition of ribosome mobility after maintaining rough microsomes at 4 degrees C after
RNase
, or antibody, treatment suggests that the ribosome binding sites are integral membrane proteins and that their mobility is controlled by the fluidity of the RER membrane. Examination of the hydrophobic interior of microsomal membranes by the freeze-fracture technique revealed the presence of homogeneously distributed 105-A intramembrane particles in control rough microsomes. However, aggregation of ribosomes by
RNase
, or their removal by treatment with puromycin, led to a redistribution of the particles into large aggregates on the cytoplasmic fracture face, leaving large particle-free regions.
...
PMID:Mobility of ribosomes bound to microsomal membranes. A freeze-etch and thin-section electron microscope study of the structure and fluidity of the rough endoplasmic reticulum. 83 67
Several procedures were used to disassemble rat liver rough microsomes (RM) into ribosomal subunits, mRNA, and ribosome-stripped membrane vesicles in order to examine the nature of the association between the mRNA of bound polysomes and the microsomal membranes. The fate of the mRNA molecules after ribosome release was determined by measuring the amount of pulse-labeled microsomal RNA in each fraction which was retained by oligo-dT cellulose or by measuring the poly A content by hybridization to radioactive poly U. It was found that ribosomal subunits and mRNA were simultaneously released from the microsomal membranes when the ribosomes were detached by: (a) treatment with puromycin in a high
salt
medium containing Mg++, (b) resuspension in a high
salt
medium lacking Mg++, and (c) chelation of Mg++ by EDTA or pyrophosphate. Poly A-containing mRNA fragments were extensively released from RM subjected to a mild treatment with
pancreatic RNase
in a medium of low ionic strength. This indicates that the 3' end of the mRNA is exposed on the outer microsomal surface and is not directly bound to the membranes. Poly A segments of bound mRNA were also accessible to [(3)H] poly U for in situ hybridization in glutaraldehyde-fixed RM. Rats were treated with drugs which inhibit translation after formation of the first peptide bonds or interfere with the initiation of protein synthesis. After these treatments inactive monomeric ribosomes, as well as ribosomes bearing mRNA, remained associated with their binding sites in microsomes prepared in media of low ionic strength. However, because there were no linkages provided by nascent chains, ribosomes, and mRNA, molecules were released from the microsomal membranes without the need of puromycin, by treatment with a high
salt
buffer containing Mg++. Thus, both in vivo and in vitro observations are consistent with a model in which mRNA does not contribute significantly to the maintenance of the interaction between bound polysomes and endoplasmic reticulum membranes in rat liver hepatocytes.
...
PMID:Release of poly A(+) messenger RNA from rat liver rough microsomes upon disassembly of bound polysomes. 88 10
The investigation of fluorescence and light-scattering change of histone F2a,
ribonuclease
, tyrosine, N-acetyltirosinamide, methyl ether tyrosine by the concentration increasing of NaCl, MgCl2, Na2SO4 in the surrounding medium was carried out. In the case of used salts the changes of tertiary structure and histones aggregations depend on the anion type, which is presented in the environment. The tertiary structure of histones formed in the presence of
salt
is stabilized by weak (hydrophobic and hydrogenic) interactions.
...
PMID:[Structure and aggregation of histones. I. Influence of the ionic composition of the medium on the structure and effectiveness of the intermolecular relationships of histone F2a (H2A+H4)]. 88
Using rice dwarf virus (RDV)-RNA which was extracted from RDV and further purified by MAK-column chromatography, anti-RNA antibodies were produced in rabbits immunized with RDV-RNA antibodies were prods immunized with RDV-RNA-methylated bovine serum albumin complexes. The antisera, as analzyed by complement fixation, cross-reacted with synthetic double stranded RNAs (poly (A)-poly (U), poly (I)-poly (C)), but not with native or denatured DNA, rRNA, tRNA, 5 S RNA and nucleic acids from rice plants. RDV-RNA treated with heat or dimethylsulfoxide was markedly reduced in reactivity to the antisera. When RDV-RNA was digested with
RNase A
at low
salt
concentration, its complement fixation activity was abolished. In double diffusion tests, two different precipitation lines were demonstrated between the antiserum and RDV-RNA. One of the precipitation lines connected with those was formed between the antisera, poly (A)-poly (U) and poly (I)-poly (C). As regards immunoglobulin classes of the antibodies, one of the two rabbits employed had antibody activity only in IgG.
...
PMID:Immunogenicity of rice dwarf virus-ribonucleic acid. 91 70
Two general methods for the isolation of DNA from various sources based on the use of cetyltrimethylammonium bromide (cetavlon, CTA-Br) are described. Cetavlon is a strong cationic detergent precipitating DNA from diluted
salt
solutions. Cells are lysed and cellular components are dissolved in the presence of cetavlon, 5 M urea, 0.1 M EDTA and 2 M NaCl (KCl). In the first method pure DNA is precipitated in the form of CTA-
salt
by direct dilution of the lysate to bring the concentration of NaCl (KCl) down to 0.5 M after the removal of the main part of proteins by deproteinization with chloroform. In the first method pure DNA is precipitated in the form of CTA-
salt
by direct dilution of the lysate to bring the concentration of NaCl (KCl) down to 0.5 M after the removal of the main part of proteins by deproteinization with chloroform. In the second method DNA is purified on the hydroxyapatite column after cell lysis and the removal of cell debris by centrifugation. Both methods are suitable for rapid isolation of pure DNA from various sources with recovery about 80% and average molecular weight 20-10(6) and higher without use of
ribonuclease
, pronase and amylase.
...
PMID:[Two simple methods for isolation of DNA from various sources using cetavlon]. 92 66
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