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Enzyme
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Query: EC:3.6.1.3 (
ATPase
)
65,361
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Purified Escherichia coli recA protein catalyzed ATP-dependent pairing of superhelical
DNA
and homologous single-stranded fragments. The product of the reaction: (i) was retained by nitrocellulose filters in 1.5 M NaCl/0.15 M Na citrate at pH 7, (ii) was dissociated at pH 12.3 but was not dissociated by heating at 55 degrees C for 4 min or by treatment with 0.2% sodium dodecyl sulfate and proteinase K, (iii) contained covalently closed circular double-stranded
DNA
(form I
DNA
), (iv) contained single-stranded fragments associated with replicative form (RF)
DNA
, and (v) contained a significant fraction of D-loops as judged by electron microscopy. Linear and nicked circular double-stranded
DNA
did not substitute well for superhelical
DNA
; intact circular single-stranded
DNA
did not substitute well for single-stranded fragments. Homologous combinations of single-stranded fragments and superhelical
DNA
from phages phiX174 and fd reacted, whereas heterologous combinations did not. The reaction required high concentrations of protein and MgCl2. The
ATPase
activity of purified recA protein was more than 98% dependent on the addition of single-stranded
DNA
. In 1 mM MgCl2, the ability of superhelical
DNA
to support the
ATPase
activity was two-thirds as good as that of single-stranded
DNA
.
...
PMID:Purified Escherichia coli recA protein catalyzes homologous pairing of superhelical DNA and single-stranded fragments. 15 61
A
DNA
-dependent
ATPase
has been purified from calf thymus. The enzyme hydrolyses ATP and dATP in the presence of heat-denatured
DNA
. It does not hydrolyse the corresponding nucleoside triphosphates of guanine, uridine and cytosine. The Km values for ATP and dATP are both 0.62 mM. The enzyme requires magnesium or manganese ions. Its sedimentation coefficient is about 4.4 S. The catalytic activity is inhibited by N-ethylmaleimide but is not sensitive to novobiocin and nalidixic acid which are potent inhibitors of bacterial DNA gyrase. In some cases, during purification, chromatographically distinct additional
DNA
-dependent
ATPase
activities were detected. Limited proteolysis or covalent modification of the enzyme in the tissues, or during the first steps of its extraction, are probably responsible for the appearance of these chromatographically distinct forms.
...
PMID:A DNA-dependent ATPase of calf-thymus. 15 29
recA protein, which is essential for general genetic recombination in Escherichia coli, promotes the homologous pairing of single-stranded
DNA
with double-stranded
DNA
to form a D loop. The amount of recA protein required for the reaction was directly proportional to the amount of single stranded
DNA
and was unaffected by similar variations in the amount of double-stranded
DNA
. The ATP analog, adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), which was not rapidly hydrolyzed by recA protein, blocked the formation of D loops but promoted the formation of stable complexes of recA protein and single-stranded
DNA
. These complexes, in turn, bound homologous or heterologous double-stranded
DNA
and partially unwound it. Because ATP gamma S competitively inhibited the
ATPase
activity of recA protein (Km/Ki approximately 300), we infer that ATP gamma S binds at a site that overlaps the site for ATP and that the functional complexes formed in the presence of the analog probably represent partial steps in the overall reaction. If the complexes formed in the presence of ATP gamma S reflect natural intermediates in the formation of D loops, recA protein must promote homologous pairing either by moving juxtaposed single-stranded and double-stranded
DNA
relative to one another or by forming and dissociating complexes reiteratively until a homologous match occurs.
...
PMID:Homologous pairing in genetic recombination: complexes of recA protein and DNA. 15 53
Two enzymes hydrolyzing ATP (
ATPase
A and
ATPase
B) were purified from freshly isolated lymphocytes of human tonsils. Both enzymes are stimulated by single-stranded
DNA
and seem to be localized in the chromatin.
ATPase
A and
ATPase
B appear to be distinct enzymes as judged from their elution profiles obtained after DEAE-cellulose and ATP-Sepharose column chromatography, from their behavior towards actinomycin D, a
DNA
intercalating agent, and from their sensitivity to monovalent salt concentration.
...
PMID:Localization of deoxyribonucleic acid-stimulated adenosine triphosphatases in human lymphocytes. 16 60
Nuclei, nuclear membranes and rough endoplasmic reticulum (rER) were isolated from onion root tips and stems. Structural preservation and purity of the fractions was determined by electron microscopic and biochemical methods. Gross compositional data (protein, phospholipid, nonpolar lipids, sterols, RNA,
DNA
), phospholipid and fatty acid patterns, enzyme activities (ATPases, ADPase, IDPase, glucose-6-phosphatase, 5'-nucleotidase, acid phosphatase, and NADH- and NADPH-cytochrome C reductases), and cytochrome contents were determined. A stable, high salt-resistant attachment of some
DNA
with the nuclear membrane was observed as well as the association of some RNA with high salt-treated nuclear and rER membranes. The phospholipid pattern was identical for both nuclear and rER membranes and showed a predominance of lecithin (about 60%) and phosphatidyl ethanolamine (20-24%). Special care was necessary to minimize lipid degradation by phospholipases during isolations. Nonpolar lipids, mostly sterols and triglycerides, accounted for 35-45% of the membrane lipids. Sterol contents were relatively high in both membrane fractions (molar ratios of sterols to phospholipids ranged from 0.12 to 0.43). Sitosterol accounted for about 80% of the total sterols. Palmitic, oleic, and linoleic acids were the most prevalent acids in membrane-bound lipids as well as in storage lipids and occurred in similar proportions in phospholipids, triglycerides and free fatty acids of the membrane. About 80% of the fatty acids in membrane phospholipids and triglycerides were unsaturated. A cytochrome of the b5 type was characterized in these membranes, but P-450-like cytochromes could not be detected. Both NADH and NADPH-cytochrome c reductases were found in nuclear and rER membranes and appeared to be enriched in rER membranes. Among the phosphatases, Mg2+-ATPase and, to lesser extents, ADPase, IDPase and acid phosphatase activities occurred in the fractions, but significant amounts of monovalent ion-stimulated
ATPase
, 5'-nucleotidase and glucose-6-phosphatase activities did not. The results obtained emphasize that the close biochemical similarities noted between rER and nuclear membranes of animal cells extend to these fractions from plant cells.
...
PMID:Characterization of nuclear membranes and endoplasmic reticulum isolated from plant tissue. 17 22
5' -Nucleotidase activity was determined in rat thyroid and some other organs employing a specific assay method. During the course of methylthiouracil (MTU) treatment, thyroid 5'-nucleotidase activity decreased significantly. This decrease was specific for this enzyme since the activity of neutral phosphatase did not change and the activity of alkaline phosphatase and Mg2+-activated
adenosine triphosphatase
increased markedly. The 5'-nucleotidase activity of the adenohypophysis also decreased following MTU treatment. This enzyme activity of the liver, heart and whole brain remained unchanged after the treatment. The role of this enzyme was discussed in relation to tissue growth and increased contents of RNA and
DNA
in the thyroid and adenohypophysis.
...
PMID:Reduction of 5'-nucleotidase activity in rat thyroid and adenohypophysis following methylthiouracil treatment. 17 98
In this first paper of a series comparing the membranes of normal lymphocyte populations from male outbred Syrian hamsters with those of neoplastic transformants (GD 248) induced by simian virus 40, a method is described for the isolation of representative plasma membrane (PM) fragments from both cell types. Multiple criteria were used to monitor the purity and yield of PM material after cell disruption by nitrogen cavitation and after membrane fractionation by a combination of differential centrifugation and isopyknic ultracentrifugation in dextran density gradients. Lactoperoxidase-catalyzed radioiodination before cell disruption was used as an extrinsic surface marker; Na+,K+-activated
ATPase
, as well as alkaline phosphatase, was used as intrinsic functional PM markers. The distribution of nuclei, mitochondria, lysosomes, and endoplasmic reticulum (ER) during fractionation was monitored by the measurement of
DNA
, succinate dehydrogenase and monoamine oxidase, beta-glucuronidase and glucose-6-phosphatase, and NADH:lipoamide oxidoreductase, respectively. According to the three PM markers employed, a 15- to 20-fold purification (over homogenate) and a PM yield of about 65% were obtained for both cell categories, with negligible contamination by
DNA
, mitochondria, lysosomes, and er. The procedure also allowed recovery of 60% of the mitochondria free of other cell elements.
...
PMID:Membranes of normal hamster lymphocytes and lymphoid cells neoplastically transformed by simian virus 40. I. High-yield purification of plasma membrane fragments. 18 92
Simian virus 40 (SV40) mRNA was isolated by hybridization of cytoplasmic RNA, from SV40-infected BS-C-1 monkey cells early in lytic infection, to SV40
DNA
immobilized on Sepharose. The early viral mRNA, when added to a wheat-germ translation system, directed the synthesis of a unique class of products including a 90,000 molecular weight (Mr) polypeptide. It was found that this 90,000 Mr product as well as a prominent 17,000 Mr polypeptide could be specifically immunoprecipitated with hamster antiserum to
SV40 T-antigen
, but not with hamster control serum. Similar immunoprecipitation of extracts of SV40-infected cells with hamster anti-T serum yielded 90,000 Mr and 17,000 Mr polypeptides; these polypeptides were not found in immunoprecipitates of uninfected cell extracts. SV40 cRNA, prepared by asymmetric transcription of plaque-purified SV40
DNA
, directed the cell-free synthesis of several products, including a 70,000 Mr polypeptide that could be specifically immunoprecipitated with anti-T serum. However, no T-antigen-related polypeptide was found in infected cells that corresponded in size to the major immunoprecipitated cRNA product.
...
PMID:Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. 19 9
The interactions between the mitochondrial and nucleocytoplasmic systems required for mitochondriogenesis have been investigated at several different levels. Those involved in the formation of functional enzyme complexes have been studied using cytochrome oxidase: this multimeric (2 X 7 and 2 X 6 subunits for enzymes from yeast and beef heart respectively) has been resolved, and the mitochondrial contribution has been shown to be dispensible for catalytic function proper. Using novel mutants, with a mitochondrial mode of inheritance, a mitochondrial gene product localized in the oligomycin-sensitive
ATPase
has been implicated in the assembly not only of this complex, but of cytochrome oxidase as well. Interactions required for the genetic competence of the mitochondrial system have become apparent as a result of studies in the mechanism of action of the highly effective mitochondrial mutagen ethidium bromide. This agent first becomes covalently inserted into mitochondrial
DNA
and, after its excision, eventually results in extensive degradation of the macromolecule. The excision reaction has now been shown to be performed by a complex between the oligomycin-sensitive
ATPase
and a DNA-binding protein presumably involved in recognizing the damage. On the level of replication and expression of the mitochondrial genome studies using thermolabile mutants have demonstrated that these processes appear independent of the replication of nuclear
DNA
but not of its expression.
...
PMID:Integration and regulation of mitochondrial assembly in yeast. 19 97
African green monkey cells (CV1 line) were infected with UV-irradiated simian virus 40 (SV40), and permissive lines of stably transformed cells were established. These cell lines display the
SV40 T-antigen
and the growth characteristics typical of nonpermissive transformed cells (e.g., reduced cell density inhibition, reduced serum dependence, ability to overgrow normal cells, and colony formation in soft agar). The level of permissiveness to superinfecting SV40 is fully comparable with that of nontransformed CV1 and BSC-1 lines. The transformed monkey lines also support SV40 plaque production under agar. By Cot analysis, the transformed permissive cells contain, on an average, 1 to 2 SV40 genome equivalents, and the majority of the viral sequences are associated with the high-molecular-weight cellular
DNA
. No spontaneous production of infectious SV40 has been observed. The transformed permissive monkey cells failed to support the replication of SV40 tsA mutants at the restrictive temperature. To account for this, it is suggested that the gene A product has separate functions for transformation and initiation of viral
DNA
synthesis, and only the former function is expressed in the transformed permissive monkey cells.
...
PMID:Properties of permissive monkey cells transformed by UV-irradiated simian virus 40. 19 53
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