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Query: EC:2.7.7.7 (
DNA polymerase
)
17,007
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
1. The
RNA-dependent DNA polymerase
associated with cytoplasmic A-type particles of murine mammary tumor virus was isolated to near homogeneity by a procedure which includes dissociation of proteins from RNA by centrifugation in a step gradient of cesium chloride, followed by an affinity chromatography on poly(rC)-agarose column. Two species of
DNA polymerase
were separated by the chromatography: enzyme I in 0.55 M NaCl and enzyme II in 0.80 M NaCl eluate, respectively. 2. The purified DNA polymerases consist of two major polypeptides, with molecular weights of 94,000 and 42,000, as the intrinsic subunits. Both enzyme protomers with a sedimentation coefficient of 6.3--6.4 S and a molecular weight of 115,000--120,000 associate to form active oligomers in low-ionic-strength buffer. 3. Both enzymes catalyzed the hydrolysis of RNA in RNA . DNA hybrids as well as the RNA-dependent synthesis of DNA; these are the intrinsic activities of the
reverse transcriptase
from B-type particles of murine mammary tumor virus as well as from avian and mammalian C-type oncornaviruses. The general catalytic properties are similar to those of the enzyme from B-type particles. Compared with DNA polymerases I,
DNA polymerase II
exhibited a high affinity for all the template-primers tested and, in addition, a high preference for (rC)N . (dG)12--18.
...
PMID:Purification and properties of RNA-dependent DNA polymerase from cytoplasmic A-type particles of murine mammary tumor virus. 8 59
A tissue culture line derived from the Asian rodent Vandeleuria oleracea has been shown to release an infectious, xenotropic type C virus. The virus-associated
reverse transcriptase
(RNA-dependent
DNA nucleotidyltransferase
) and the major internal protein p30 are immunologically related to the respective proteins of the woolly monkey-gibbon ape group of infectious primate viruses. By these criteria the V. oleracea viral isolate is similar to the murine type C-I class of endogenous retroviruses and has been designated Vand C-I. Nucleic acid homology studies show that V. oleracea cellular DNA shares similar levels of homology with DNA from members of the Mus and Rattus genera and lower levels of homology with other rodent genera. The Vand C-I viral genome is present in V. oleracea cellular DNA in multiple copies, and partially related sequences can be detected in other rodent genera. These results support the conclusion that the Vand C-I viral genome is genetically transmitted in V. oleracea and that the type C-I class of endogenous retroviral genes has been highly conserved during evolution.
...
PMID:Isolation of an endogenous type C virus related to the infectious primate type C viruses from the Asian rodent Vandeleuria oleracea. 9 Jan 55
An
RNA-directed DNA polymerase
was purified from a cell line derived from a radiation-induced lymphoma in NIH Swiss mice which produced non-infectious type C virus particles. The enzyme was isolated from a high speed particulate fraction which bands at a density of 1.16--1.19 g/ml in a sucrose gradient, and purified by successive chromatography on DEAE-cellulose, phosphocellulose and hydroxyapatite. The purified
DNA polymerase
has a molecular weight of 68 000, a pH optimum of 7.5, a KCl optimum of 50 mM, and a Mn2+ optimum of 0.25 mM. It prefers (dT)15 . (A)n to (dT)15 . (dA)n as the primer template and transcribes the poly(C) strand of (dG)15 .(C)n and (dG)15 . (OMeC)n. It transcribes heteropolymeric regions of avian myeloblastosis virus 70 S RNA, and is inhibited by antiserum to Rauscher murine leukemia virus
DNA polymerase
. Comparison of the properties of
DNA polymerase
purified from radiation-induced lymphoma cells with the
DNA polymerase
purified from non-defective murine type C RNA tumor viruses shows that the mouse lymphoma enzyme is both biochemically and immunologically related to murine leukemia virus DNA polymerases.
...
PMID:Characterization of an RNA-directed DNA-polymerase from a cell line derived from a radiation-induced lymphoma in mice. 9 May 22
A complex between tRNATrp (beef) and 35 S RNA from avian myeloblastosis virus is obtained when the mixture is preincubated in the presence of
reverse transcriptase
at 35 degrees C. The tRNA-RNA complex is active in initiating DNA synthesis catalyzed by
reverse transcriptase
. The interaction of tRNA with
reverse transcriptase
involves the partial unwinding of the acceptor stem of tRNA, as evidenced by nuclease digestion with RNAase T1 and micrococcal nuclease. When tRNA2Glu (coli), having a high degree of similarity with primer tRNA at the level of the acceptor stem, was used as primer for DNA synthesis, a low but significant level of incorporation was obtained, if the reaction was performed at 35 degrees C, while a high incorporation, similar to the one obtained with tRNATrp was obtained when the annealing between tRNA2Glu and 35 S RNA was performed at 80 degrees C. Our evidences point out to an important role of the viral
DNA polymerase
in positioning the primer on the RNA genome.
...
PMID:Reverse transcriptase mediated binding of primer tRNA to the viral genome. 9 Nov 58
A restriction fragment strand complementary to a sequence near the 3' end of Escherichia coli 16S rRNA has been used to prime
reverse transcriptase
(avian myeloblastosis virus RNA-directed
DNA nucleotidyltransferase
; deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase,
EC 2.7.7.7
). In addition to transcripts that were extended to the 5' end of the RNA, two major transcription intermediates were observed. These discrete-sized cDNA intermediates are the result of a kinetic barrier imposed by monomethylation of the amino group on guanine that participates in base-pairing. Both major transcription intermediates correspond to attenuation at the known positions of N2-methylguanine (m2G) in the rRNA sequence. The relaxation time for elongation of the cDNA through m2G is approximately 3 min. No other major kinetic pauses were observed in the 1340 bases transcribed.
...
PMID:Reverse transcriptase pauses at N2-methylguanine during in vitro transcription of Escherichia coli 16S ribosomal RNA. 9 Nov 69
Benzophenanthridine alkaloids, fagaronine 4, O-methylfagaronine 5, nitidine 1, allonitidine 3 and methoxydihydronitidine 2 have been shown to posses inhibitory activity against
reverse transcriptase
of RNA tumor viruses. The enzyme inhibition (50%) by these alkaloids was found in the range of 6-60 microgram per milliliter of the reaction mixture when polynucleotide-oligodeoxynucleotide complexes were used as template primers. The results suggested that the benzophenanthridine alkaloids interacted with the template primers (particularly of the A:T base pairs) and not with the enzyme proteins. Kinetics reaction of the reverse transciptase inhibition showed that the alkaloids stopped the
DNA polymerase
synthesis instantly, probably by interacting with the template primer.
...
PMID:Inhibition of reverse transcriptase activity by benzophenanthridine alkaloids. 9 65
Two mutants of avian sarcoma virus which exhibit different phenotypes have been analyzed for the properties of their
RNA-dependent DNA polymerase
and RNase H activities. LA 338 is a complex multiple mutant with at least one lesioneach in transformation and replication functions. The purified
RNA-dependent DNA polymerase
-RNase H complex from the mutant is twofold more thermolabile than that from the wild-type parent. A peculiarity of this mutant is that the ability of the enzyme to respond to synthetic template-primers is lost more rapidly than is the response to native RNA as template. The mutant enzyme cannot be protected from inactivation by the addition of synthetic template-primers. LA 672 represents a different phenotype among reverse transciptase mutant, showing a "late"-acting block in replication which affects only production of progeny by infected cells grown at the nonpermissive temperature. The purified
DNA polymerase
-RNase H complex of LA 672 is not thermolabile; rather, progeny grown at the nonpermissive temperature yield purified enzyme with a 20-fold-reduced specific activity in both
DNA polymerase
and RNase H. The content of
reverse transcriptase
protein in such noninfectious progeny, furthermore, did not appear to be significantly diminished since immunologically active enzyme could be demonstrated in a competition test for anti-
reverse transcriptase
antibody and since beta and alpha subunits of
reverse transcriptase
could be identified after polyacrylamide gel electrophoresis of partially purified enzyme preparations. The amounts of beta and alpha from the mutant were about twofold lower.
...
PMID:Two avian sarcoma virus mutants with defects in the DNA polymerase-RNase H complex. 9 85
The
reverse transcriptase
(RNA-dependent
DNA nucleotidyltransferase
) of the type C RNA virus produced by the human lymphoma cell line SU-DHL-1 was purified by ion-exchange chromatography of SU-DHL-1 culture fluids and repetitive affinity chromatography on poly(rC).agarose, as were the polymerases of several other type C viruses. The DHL-1 enzyme used template-primers at levels expected of a viral
reverse transcriptase
, and sodium dodecyl sulfate gel electrophoretic analysis of radioiodinated DHL-1 enzyme revealed a peak at a position corresponding to those of several other type C viral reverse transcriptases (namely, at 72,000-78,000 daltons). The purified enzyme was partially neutralized by antibodies specific for the
reverse transcriptase
of simian sarcoma virus. Two-dimensional analysis on thin-layer cellulose plates of tryptic hydrolysates of the radioiodinated enzymes of several viruses revealed that six peptides are common to the polymerases of simian sarcoma virus, gibbon ape leukemia virus, baboon endogenous virus, and the DHL-1 virus, and that two to four peptides are unique to each of these enzymes. The DHL-1 viral
reverse transcriptase
appears to be most closely related structurally to the enzymes of simian sarcoma virus, gibbon ape leukemia virus, and baboon endogenous virus. However, the DHL-1 viral enzyme differed from any one or combination of the other subhuman primate viral enzymes by virtue of its unique peptides. The implications of these findings with respect to the probable origin of the DHL-1 virus are discussed.
...
PMID:Characterization of the reverse transcriptase of a type C RNA virus produced by a human lymphoma cell line. 9 23
RNA-directed DNA polymerase
was isolated from the liver, spleen, and myeloblasts of chickens that had been infected with virus of avian myeloblastosis. The enzyme was chromatographically purified from the myeloblasts and brought to about 1,000-fold concentration. The method consisted in cell fractionation, lysis of the microsomal fraction, chromatography on Sephadex G-200 and phosphocellulose, as well as ultracentrifugation in the glycerol gradient. The cellular DNA polymerases alpha and beta were clearly separated from the
DNA polymerase
of avian myeloblastosis virus and could be distinguished from one another by template-specific reactions. The viral
DNA polymerase
activities in the microsomal fractions of liver, spleen, and myeloblasts were compared with one another. The amount of avian myeloblastosis virus and related
DNA polymerase
recorded from the myeloblasts was about six times that in the liver and four times higher than that in the spleen. The procedure described, together with the use of cell fractionation and gel filtration, is an appropriate method for biochemical detection of avian oncorna viruses in cells.
...
PMID:[Separation of cellular and viral DNA polymerase from oncornavirus infected chicken cells]. 9 56
Hepatitis B virus DNA made fully double stranded by a virion
DNA polymerase
reaction could be converted from circular to linear molecules by heating in 10 mM NaCl at 77 degrees C or in 100 mM NaCl at 90 degrees C for 15 min. Heat-generated linear hepatitis B virus DNA was reannealed to circular molecules by incubating in higher salt concentrations. The identity of the molecular forms was established by their electrophoretic mobility and appearance in electron micrographs. Recircularization was blocked by reacting linear molecules with nuclease S1 or avian myeloblastosis virus
reverse transcriptase
. These results suggest that the heated linear DNA had single-stranded ends with complementary nucleotide sequences. It also suggests that a discontinuity or nick is present in each strand of the circular DNA molecule after the single-stranded region is made double stranded by the virion
DNA polymerase
reaction. The difference in contour length by electron microscopy of circular and linear molecules spread under aqueous conditions suggested that the discontinuities in the two strands were about 270 base pairs apart. The amount of nucleotide incorporated into the ends of heat-generated linear hepatitis B virus DNA by
reverse transcriptase
suggested that the single-stranded ends were about 305 bases in length. This fully double-stranded linear DNA was cleaved with EcoRI or HpaI restriction endonuclease. The sum of the two fragments generated by each totaled 3,510 base pairs, 310 base pairs greater than the contour length of circular hepatitis B virus DNA which represents a third estimate of the distance between the discontinuities in the two DNA strands of circular DNA. Restriction endonuclease cleavage also indicated that the ends of heated linear DNA which correspond to the discontinuities in the two strands of the circular DNA are at unique sites in the DNA with respect to the restriction sites.
...
PMID:Hepatitis B viral DNA molecules have cohesive ends. 9 58
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