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At least 937 records · Page 52Linked to original sources
PubMed1981

Induction of immunoenhancing factors for murine splenocyte cultures by Salmonella typhimurium ribosome and ribonucleic acid extracts.

Ribosomal and ribonucleic acid (RNA)-rich preparations derived from Salmonella typhimurium were examined for their ability to enhance the primary in vitro antibody response of normal mouse spleen cell cultures to sheep erythrocytes. Both of these fractions were consistently more active in elevating the antibody response of normal mouse splenocytes from lipopolysaccharide (LPS) responder mice than was LPS. Furthermore, injection of mice with either the ribosomal or RNA-rich fraction induced antibody response helper factor activity in 2-h post-treatment serum similar to that induced by LPS. Endotoxin low-responding C3H/HeJ mice were stimulated to release helper factors by ribosomes and the RNA extracts but not by LPS. Treatment of the ribosomes and RNA fractions with ribonuclease destroyed their ability to stimulate the production of the helper factor in serum of treated mice. Therefore, it appears likely that ribosomes and RNA-rich fractions stimulated an intermediate helper factor due to the presence of RNA and not LPS.

Animals↗
PubMed1975

Isolation and partial characterization of three Escherichia coli mutants with altered transfer ribonucleic acid methylases.

Seven transfer ribonucleic acid (tRNA) methylase mutants were isolated from Escherichia coli K-12 by examining the ability of RNA prepared from clones of unselected mutagenized cells to accept methyl groups from S-adenosylmethionine catalyzed by crude enzymes from wild-type cells. Five of the mutants had an altered uracil-tRNA methylase; consequently their tRNA's lacked ribothymidine. One mutant had tRNA deficient in 7-methylguanosine, and one mutant contained tRNA lacking 2-thio-5-methylaminomethyluridine. The genetic loci of the three tRNA methylase mutants were distributed over the E. coli genome. The mutant strain deficient in 7-methylguanosine biosynthesis showed a reduced efficiency in the suppression of amber mutations carried by T4 or lambda phages.

Chromatography, Affinity↗
PubMed1977

Characterization and in vitro translation of polyadenylated messenger ribonucleic acid from Neurospora crassa.

Ribonucleic acid (RNA) extracted from Neurospora crassa has been fractionated by oligodeoxythymidylic acid [oligo(dT)]-cellulose chromatography into polyadenylated messenger RNA [poly(A) mRNA] and unbound RNA. The poly(A) mRNA, which comprises approximately 1.7% of the total cellular RNA, was further characterized by Sepharose 4B chromatography and polyacrylamide gel electrophoresis. Both techniques showed that the poly(A) mRNA was heterodisperse in size, with an average molecular weight similar to that of 17S ribosomal RNA (rRNA). The poly(A) segments isolated from the poly(A) mRNA were relatively short, with three major size classes of 30, 55, and 70 nucleotides. Gel electrophoresis of the non-poly(A) RNA indicated that it contained primarily rRNA and 4S RNA. The optimal conditions were determined for the translation of Neurospora mRNA in a cell-free wheat germ protein-synthesizing system. Poly(A) mRNA stimulated the incorporation of [14C]leucine into polypeptides ranging in size from 10,000 to 100,000 daltons. The RNA that did not bind to oligo(dT)-cellulose also stimulated the incorporation of [14C]leucine, indicating that this fraction contains a significant concentration of mRNA which has either no poly(A) or very short poly(A) segments. In addition, the translation of both poly(A) mRNA and unbound mRNA was inhibited by 7-methylguanosine-5'-monophosphate (m7G5'p). This is preliminary evidence for the existence of a 5'-RNA "cap" on Neurospora mRNA.

Chromatography, Agarose↗
PubMed1979

Escherichia coli mutant strain with altered expression of the tryptophan operon: ribonucleic acid synthesis in vitro.

Ribonucleic acid (RNA) synthesis has been studied in vitro with a partially purified preparation of RNA polymerase from a mutant strain of Escherichia coli with a reduced rate of accumulation of tryptophan RNA (P.H. Pouwels and H.J. Scholten, J. Bacteriol. 139:393-397, 1979). The incorporation of radioactive label into RNA with polymerase from mutant bacteria is considerably lower than that with the enzyme from wild-type bacteria. These results are explained by the presence in mutant bacteria, but not in wild-type bacteria, of a factor which suppresses the accumulation of RNA. Mutant bacteria contain a factor which renders RNA synthesis with mutant and wild-type RNA polymerase resistant to various inhibitors of RNA synthesis, e.g. rifampin, streptolydigin, and heparin. We conclude that in mutant bacteria a factor is modified which suppresses the accumulation of some RNA species and lowers the sensitivity of RNA polymerase to some transcription inhibitors.

Aminoglycosides↗
PubMed1972

Development of bacteriophage alpha. II. Dependence on the host ribonucleic acid polymerase.

Measurements of ribonucleic acid (RNA) synthesis using (14)C-uracil uptake in rifampin-sensitive and rifampin-resistant strains of Bacillus megatherium were carried out after infection of the bacteria with bacteriophage alpha. Phage development was inhibited in the former but not the latter strain, showing that alpha phage development is dependent on the bacterial host RNA polymerase. This dependence exists at all times during eclipse. RNA polymerase extracts showed the same in vitro rifampin sensitivity as the corresponding bacterial strains.

Bacillus megaterium↗
PubMed1972

Ribonuclease H: a ubiquitous activity in virions of ribonucleic acid tumor viruses.

Ten ribonucleic acid (RNA) tumor viruses grown in five different host cell species and three non-oncogenic viruses from three different virus groups have been examined for ribonuclease H content. Three different substrates were used to assay ribonuclease H: calf thymus [(3)H]RNA-deoxyribonucleic acid (DNA) hybrid prepared with denatured calf thymus DNA and Escherichia coli DNA-directed RNA polymerase, (3)H-polydenylic acid [(3)H-poly(A)] complexed to polydeoxythymidylic acid [poly(dT)], and (3)H-polyuridylic acid [(3)H-poly(U)] complexed to polydeoxyadenylic acid [poly(dA)]. All ten RNA tumor viruses contained ribonuclease H activity which degraded the RNA of both the calf thymus hybrid and poly(A)-poly(dT), whereas only the ribonuclease H in the Moloney strain of murine sarcoma-leukemia virus and in RD-feline leukemia virus hydrolyzed the RNA strand of poly(U)-poly(dA). No appreciable ribonuclease H activity was detected in influenza, Sendai, or vesicular stomatitis virus. The ribonuclease H and RNA-directed DNA polymerase activities in Moloney murine sarcoma-leukemia virus were inseparable by phosphocellulose chromatography or glycerol gradient centrifugation, but appeared to be partially separated by diethylaminoethyl-cellulose chromatography.

Animals↗
PubMed1972

Transcription during the development of bacteriophage phi 29: production of host- and phi 29-specific ribonucleic acid.

The synthesis of ribonucleic acid (RNA) during development of the virulent Bacillus subtilis bacteriophage phi29 has been analyzed. Transcription of host deoxyribonucleic acid (DNA) continues at the preinfection rate throughout the latent period of viral growth. RNA-DNA hybridization was used to show that host messenger RNA synthesis continues late into the phage lytic cycle. Amino acid-labeling experiments show that this RNA is continuously used to produce protein. Ribosomal RNA production is not inhibited by phage infection. Small quantities of phage-specific RNA first appear between min 6 and 9 after infection. This RNA is made exclusively from one of the phi29 DNA strands. At 12 min postinfection, when phage DNA replication commences, large quantities of viral RNA start to be synthesized. This RNA appears to be transcribed from both strands of phi29 DNA. Studies with rifamycin and rifamycin-resistant host strains showed that the production of all phage phi29-specific RNA requires those components of the host RNA polymerase which are sensitive to this antibiotic. Thus, phage phi29 does not stop transcription of host DNA and may produce only one element for regulation of transcription of its own DNA. These findings may reflect the limited amount of genetic information carried by this phage.

Autoradiography↗
PubMed1973

Transcription during the development of bacteriophage phi29: definition of "early" and "late" phi29 ribonucleic acid.

Bacteriophage phi29 messenger ribonucleic acid (mRNA) production following infection of Bacillus subtilis has been analyzed. Early (e) phi29 RNA, made prior to the onset of phage deoxyribonucleic acid (DNA) replication and exclusively from the light (L) phi29 DNA strand, has been shown by RNA-DNA hybridization-competition experiments to be present throughout the phage latent period. No repression of e RNA production during phi29 development could be demonstrated. Unmodified host RNA polymerase molecules appear to be sufficient for the synthesis of e RNA since phage-specific RNA made in the presence of chloramphenicol hybridizes only to the L strand of phi29 DNA, and this RNA can be effectively competed during hybridization by e RNA. The appearance of late (l) phi29 RNA is coincident with the onset of viral DNA replication. This RNA consists of L DNA strand transcripts which are identical to e RNA and a new class of mRNAs made exclusively from the "heavy" (H) phi29 DNA strand (lH). Protein synthesis in infected cells is required for lH RNA production. Studies with the antibiotic rifamycin demonstrated that synthesis of the major phi29 structural proteins is dependent on production of lH RNA.

Amino Acids↗
PubMed1968

Induction of poxvirus ribonucleic acid polymerases.

Two distinct ribonucleic acid polymerase activities were induced in HeLa cells by poxvirus infection. These activities differ both in their properties and the time of their appearance after infection. One catalyzes the dAT (copolymer of deoxyadenylate and deoxythymidylate)-primed conversion of adenosine triphosphate and uridine triphosphate into an acid-insoluble product. This enzyme is detectable only if deoxyribonucleic acid synthesis has been blocked. In contrast, the accumulation of progeny genomes is a necessary condition for induction of the second enzyme. The latter activity, which is unmasked by detergent treatment, is found exclusively in maturing virus particles. The possibility that both enzymes are involved in transcribing the viral genome is discussed.

Cycloheximide↗
PubMed1971

Virus protein synthesis in animal cell-free systems: nature of the products synthesized in resonse to ribonucleic acid of encephalomyocarditis virus.

Ribonucleic acid (RNA) from encephalomyocarditis (EMC) virus stimulates the incorporation of amino acids into protein in cell-free protein-synthetic systems derived from Krebs mouse ascites tumor cells and chick embryo fibroblasts; the mouse system is the more responsive to the viral RNA. The greater part of this difference in activity can be ascribed to the cell sap, but the origin of the ribosomes also has a marked effect. The nature of the polypeptides formed in these cell-free systems was investigated by electrophoresis on polyacrylamide gels and by fingerprint analysis of tryptic digests. The same product in part appears to be synthesized in response to the EMC RNA in both systems. It was not detected if the EMC RNA was partly degraded (</=4S) or replaced by other species of RNA, including that from influenza virus. The results suggest that EMC RNA is partially translated in these systems to yield virus-specific polypeptides.

Amino Acids↗
PubMed1971

Electron microscope study of ribonucleic acid of myxoviruses.

Intact ribonucleic acid (RNA) molecules in an extended form were extracted from purified influenza virus and observed in the electron microscope. For this study, the RNA extraction procedure and the Kleinschmidt protein monolayer technique were modified. The mean lengths of RNA from X7, X7-F1, and WSN strains of influenza virus were found to be 2.69, 2.55, and 2.37 mum, respectively. From these measurements, the corresponding estimated molecular weights would be 2.9, 2.8, and 2.5 x 10(6) daltons. X7 and WSN RNA preparations were exposed to pH 3 to disrupt intact molecules. Histograms of length measurements showed five peaks, which were interpreted to represent the five pieces of RNA reported to exist in the influenza virion. X7 RNA appeared to be more stable than WSN RNA when stored at 4 C. The profiles of histograms of incomplete virus RNA suggest that the high molecular-weight component is missing. In preliminary experiments on Newcastle disease virus RNA, molecules of various lengths were observed.

Centrifugation, Density Gradient↗
PubMed1971

Virus interference by cellular double-stranded ribonucleic acid.

Ribonuclease-resistant ribonucleic acid (RNA) was isolated from uridine-labeled cultures of rabbit kidney, chicken embryo, and HeLa cells. This RNA, regardless of its source, was found to induce interference with virus growth in either rabbit kidney or chicken embryo cultures. Nuclease-treated cellular nucleic acids exhibited interference-inducing activity which eluted with a small fraction of RNA in the exclusion volume of a 6% agarose gel column. Besides resistance to ribonucleases, the interference inducer and RNA isolated from partially digested nucleic acids have in common two properties of double-stranded RNA: (i) similar sharp melting profiles were obtained for inducer and ribonuclease-resistant RNA, with T(m) dependent on NaCl concentration; (ii) ribonuclease-resistant inducer and RNA banded together in Cs(2)SO(4) density gradients at a density characteristic of known double-stranded RNA. After melting at low ionic strength, the labeled RNA shifted to a higher density and its capacity to inhibit virus replication was lost. Velocity sedimentation analysis of the cellular ribonuclease-resistant RNA indicated that the majority sedimented between 7 and 11S, but only RNA sedimenting at >==8 to 20S had a high specific activity of interference induction. Without prior ribonuclease treatment, the ribonuclease-resistant RNA can be precipitated with 2 m LiCl and thus appears to exist in purified cellular nucleic acids as part of molecular complexes with both single- and double-stranded regions of RNA. The biosynthesis of cellular double-stranded RNA is inhibited by actinomycin D.

Animals↗
PubMed1995

Changes in estrogen receptor messenger ribonucleic acid in sheep fetal and maternal tissues during late gestation and labor.

OBJECTIVE: Our purpose was to investigate whether there is an increase in messenger ribonucleic acid for estrogen receptor in critical maternal or fetal tissues in the last third of pregnancy and during labor in sheep. STUDY DESIGN: Estrogen receptor messenger ribonucleic acid was measured by Northern hybridization analysis in fetal-placental and maternal uterine tissues during the last third of pregnancy and during spontaneous or cortisol-induced labor in sheep. Statistical differences were assessed with two-way analysis of variance. RESULTS: No estrogen receptor messenger ribonucleic acid was observed in amnion or chorion in any animal studied. There were no gestational age related changes in estrogen receptor messenger ribonucleic acid in any tissues between 100 and 145 days' gestation. During spontaneous and cortisol-induced labor estrogen receptor messenger ribonucleic acid increased significantly (p < 0.05) in myometrium, endometrium, and cervix. No increase was observed in the fetal placental cotyledon and mesometrium. Estrogen receptor messenger ribonucleic acid was markedly decreased (p < 0.05) in myometrium and endometrium after fetal adrenalectomy. CONCLUSION: An increase in estrogen receptor messenger ribonucleic acid in association with labor may contribute part of the mechanism by which estrogens exert their influence on the process of parturition.

Animals↗
PubMed1993

Changes in gap junction connexin-43 messenger ribonucleic acid levels associated with rat ovarian follicular development as demonstrated by in situ hybridization.

OBJECTIVE: The purpose was to evaluate the changes in gap junction connexin-43 messenger ribonucleic acid levels associated with rat ovarian follicular development. Gap junctions connect the plasma membranes of adjacent cells through cell-to-cell channels, allowing synchronization of cellular events, including ovarian follicular development. Ovarian gap junctions consist of the protein connexin-43. STUDY DESIGN: We used the hypophysectomized immature rat treated with estrogen or gonadotropins as a model to study the ovarian regulation of connexin-43 messenger ribonucleic acid. In situ hybridization with radiolabeled riboprobes was used to localize and quantitate connexin-43 messenger ribonucleic acid. RESULTS: We demonstrated that connexin-43 messenger ribonucleic acid was localized to follicular granulosa cells. Estrogen significantly up-regulated connexin-43 messenger ribonucleic acid (91%), whereas gonadotropins that stimulate ovulation and corpus luteum formation completely down-regulated the connexin-43 gene. These results correlate closely with previous immunohistochemical studies of connexin-43 protein. CONCLUSION: The positive correlation between follicular development and granulosa cell content of connexin-43 messenger ribonucleic acid is caused by transcriptional activation of the gap junction connexin-43 gene, posttranscriptional stability of connexin-43 messenger ribonucleic acid, or both. Future studies will determine the molecular mechanisms of hormonal regulation of the connexin-43 gene.

Animals↗
PubMed1975

The urinary excretion of nucleosides of ribonucleic acid by patients with advanced cancer.

By means of a sensitive and specific method utilizing gas-liquid chromatography, the excretion levels for three nucleosides, degradation minor base products of ribonucleic acid, primarily transfer ribonucleic acid, were determined in 24-hour urine specimens from over 200 patients with solid tumor malignancies. These nucleosides were N2,N2-dimethylguanosine, l-methylinosine, and pseudouridine. When compared to normal control values, elevated levels of these compounds were found for patients in each of several tumor types studied. Increases in pseudouridine excretion suggest increased tumor transfer ribonucleic acid turnover; in addition, for the methylated nucleosides, higher than normal values may reflect enhanced transfer ribonucleic acid methylase activity of the neoplastic cells.

Chromatography, Gas↗
PubMed1997

Determination of degradation rates of transfer and ribosomal ribonucleic acids in cultured rat hepatocytes by measuring N6-threoninocarbonyladenosine, dihydrouridine, and pseudouridine in medium using high-performance liquid chromatography.

Modified ribonucleic acid catabolites excreted into the medium by primary cultures of rat hepatocytes (2.3 +/- 0.42 x 10(6) cells/dish) during a 24-h cultivation period were quantified by reversed-phase high-performance liquid chromatography (fmol/10(3) cells): 613 +/- 81 dihydrouridine, 46 +/- 6 N6-threoninocarbonyladenosine, 1879 +/- 220 pseudouridine. On the basis of these excretion rates and the average frequency of occurrence of these modified ribonucleosides per cytoplasmic transfer ribonucleic acid (residues: 2.6 dihydrouridine, 0.22 N6-threoninocarbonyladenosine, 3 pseudouridine) as well as per cytoplasmic ribosomal ribonucleic acid (residues: 95 pseudouridine), the degradation rates of transfer and ribosomal ribonucleic acids were calculated. The degradation rate of transfer ribonucleic acid (fmol/10(3) cells/24 h) was 236 +/- 31 (via dihydrouridine) and 211 +/- 28 (via N6-threoninocarbonyladenosine) and that of ribosomal ribonucleic acid (fmol/10(3) cells/24 h) was 13.1 +/- 1.7 (via pseudouridine and N6-threoninocarbonyladenosine).

Adenosine↗
PubMed1996

Decreased very-low-density lipoprotein and low-density lipoprotein receptor messenger ribonucleic acid expression in placentas from preeclamptic pregnancies.

OBJECTIVE: Our purpose was to investigate the levels of very-low-density lipoprotein and low-density lipoprotein receptor messenger ribonucleic acid expression in placentas obtained from normal pregnant women at various gestational stages and from patients with preeclampsia. STUDY DESIGN: The relative level of expression of very-low-density lipoprotein and low-density lipoprotein receptor messenger ribonucleic acid in each sample was determined by Northern blot analysis as a ratio of the intensity to that of glyceraldehyde-3-phosphate dehydrogenase messenger ribonucleic acid. RESULTS: The expression of very-low-density lipoprotein receptor messenger ribonucleic acid increased significantly (p < 0.05) from both the first (n = 9) and second (n = 8) trimesters to the third (n = 11) trimester. Similarly, the expression of low-density lipoprotein receptor messenger ribonucleic acid increased significantly (p < 0.05) from the first to the third trimester. The expression of both very-low-density lipoprotein and low-density lipoprotein receptor messenger ribonucleic acids in third-trimester placentas from preeclamptic pregnancies (n = 13) was significantly lower than that in normal pregnancies in the third trimester. CONCLUSION: These results demonstrate the expression of very-low-density lipoprotein and low-density lipoprotein receptor messenger ribonucleic acids in the human placenta, both of which increase at late gestational stages, and the decreased expression in cases of preeclampsia. Abnormal fetomaternal lipid metabolism resulting from low expression of these receptors in the placenta may be involved in the pathogenesis of preeclampsia.

Adult↗
PubMed1995

Insulin and glucose modulate glucose transporter messenger ribonucleic acid expression and glucose uptake in trophoblasts isolated from first-trimester chorionic villi.

OBJECTIVE: Our purpose was to determine the effects of insulin and glucose on glucose transport and expression of GLUT1 glucose transporter messenger ribonucleic acid in first-trimester human trophoblast-like cells. STUDY DESIGN: First-trimester human trophoblast-like cells were maintained as a continuous cell line. For 2[3H]deoxy-D-glucose uptake and messenger ribonucleic acid studies the cells were incubated in the presence or absence of insulin (10(-7) to 10(-11) mol/L) or D-glucose (0 to 50 mmol/L) for 0 to 24 hours. Glucose transport was measured by incubating cells with 0.1 mmol/L 2[3H]deoxy-D-glucose for 5 minutes. Specific uptake was determined by incubating companion cultures with 10 mumol/L cytochalasin B. The cells were then solubilized with sodium hydroxide and the radioactivity counted. Data were expressed as nanomoles of 2[3H]deoxy-D-glucose transported per milligram of protein per 5 minutes and analyzed by one-way analysis of variance with post hoc testing by the method of Tukey. GLUT1 messenger ribonucleic acid was measured by Northern blotting of total ribonucleic acid samples hybridized to a phosphorus 32-labeled complementary deoxyribonucleic encoding the rat GLUT1 glucose transporter. As a control for loading efficiency, blots were stripped and rehybridized to a 40-mer phosphorus 32-labeled beta-actin oligonucleotide probe. RESULTS: Insulin treatment resulted in a dose-dependent increase in the transport of 2[3H]deoxy-D-glucose at 24 hours (p < 0.001 at 10(-7) mol/L). This change was first detected at 12 hours of incubation. These data closely paralleled the insulin-induced increase in GLUT1 messenger ribonucleic acid seen in Northern blots. In contrast to insulin, increasing concentrations of D-glucose did not change the transport of 2[3H]deoxy-D-glucose. However, when cells were incubated in low concentrations of D-glucose (0 or 1 mmol/L), an enhancement in the uptake of 2[3H]deoxy-D-glucose (p < 0.001) was observed. Kinetic studies indicated that D-glucose augmentation of 2[3H]eoxy-D-glucose uptake was significant at 9 hours (p < 0.05). The effects of D-glucose on GLUT1 messenger ribonucleic acid expression paralleled the uptake of 2[3H]deoxy-D-glucose, although the modulation of GLUT1 messenger ribonucleic acid levels by glucose was much less pronounced than in insulin-treated cells. CONCLUSION: Although it has been assumed that the placenta has a limited role in influencing glucose transport to the fetus, our in vitro data demonstrate that both insulin and glucose can modulate glucose transport at the cellular level of the placental trophoblast. Thus maternal insulin and glycemic status may influence the expression of GLUT1, the major trophoblast glucose transporter protein, therefore directly affecting first-trimester placental glucose transport. These in vitro data may help explain the association between maternal glucose abnormalities and impaired fetal development during the first trimester when placental GLUT1 messenger ribonucleic acid expression is at its peak.

Analysis of Variance↗