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At least 19 recordsLinked to original sources
PubMed1982

Detection of RNA complementary to herpes-simplex virus in mononuclear cells from patients with Behçet's syndrome and recurrent oral ulcers.

Viral DNA probes were used in in-situ hybridisation to detect the complementary RNA in mononuclear cells from the blood of normal subjects and patients with Behçet's syndrome and recurrent oral ulcers. Hybridisation of 125I-labelled DNA probes was detected by means of autoradiography and quantitated with a video image-analysis technique. Hybridisation between herpes-simplex virus type 1 (HSV-1) DNA and the complementary RNA in mononuclear cells was significantly greater in 10 of 20 patients with Behçet's syndrome than in controls. Consistently, mononuclear cells from fewer patients showed significant hybridisation with the HSV-2 probe, and the grain counts were also lower. Control DNA probes from adenovirus 2, bacteriophage lambda, and Micrococcus lysodeikticus did not show significant hybridisation. Further separation of the patients into the four types of Behçet's syndrome revealed that mononuclear cells in 8 of 10 patients with the ocular or arthritic types showed significant hybridisation of RNA to the HSV-1 DNA probe, compared with 2 of 10 patients with the mucocutaneous or neurological types. Mononuclear cells from 4 of 8 patients with minor but not major aphthous ulcers also showed significant RNA hybridisation to HSV-1 DNA. The results suggest that at least part of the HSV genome is present and transcribed in peripheral-blood mononuclear cells--probably lymphocytes--of patients with the ocular and arthritic types of Behçet's syndrome and minor aphthous ulcers. The immunopathogenesis of these diseases might therefore be associated with HSV-1.

Behcet Syndrome↗
PubMed1990

An antibody directed against a peptide encoded by RNA complementary to mRNA for vasopressin recognizes putative vasopressin receptors.

The purpose of this study was to generate an antibody against the binding site of the vasopressin receptor. Recently it has been demonstrated that complementary RNA sequences may encode interacting peptides. We determined whether or not an antibody directed against a peptide (PVA) specified by RNA complementary to the mRNA of rat arginine vasopressin (AVP) would recognize the AVP receptor. The antibody was purified sequentially over a protein A and then a PVA affinity column. A specific anti-PVA antibody affinity column also was made. The specific anti-PVA antibody recognized two proteins with molecular weights of 76 and 70 kD in homogenates from kidney and brain tissue containing the hypothalamus/thalamus/septum. This antibody also blocked binding of 3H-AVP to primary cultured neuronal cells, whereas the nonspecific antibody did not. Furthermore, the blocking efficiency of the antibody increased when more of the specific anti-PVA antibody was present. We also determined whether or not the antiserum contained any biological activity by observing urine volume and urine osmolality in the presence or absence of preimmune or immune serum. Only the immune serum was able to reverse the antidiuretic and urine-concentrating effects of AVP, suggesting that the antibody antagonized the effects of AVP on the kidney. Taken together with the observation that this antibody did not bind to vasopressin, these data strongly indicate that an antibody to the vasopressin receptor binding site has been made and add further support to the molecular recognition theory.

Amino Acid Sequence↗
PubMed1985

[Analysis of ribonucleoproteins of influenza virus containing genomic and complementary RNA].

The density and sedimentation characteristics of ribonucleoproteins (RNP) containing genomic RNA from influenza virus and RNA complementary have been studied. Radioactive RNA from infected cells has been used for analysis. RNA classes of interest were isolated by reannealing with abundant nonradioactive genomic and complementary RNA and separation of resulting duplexes in electrophoresis. The RNP containing antigenomic virus-specific RNA are practically identical to "genomic" RNP for their sedimentation and density characteristics. The "plus" RNP is characterized by the stoichiometric mode of RNA protein interaction.

Animals↗
PubMed1993

Studies on the peptides encoded by rat and human angiotensin II complementary RNA.

Some evidence suggests that RNA complementary to the messenger RNA encoding a peptide hormone encodes a complementary peptide that binds the original peptide hormone. The objective of this investigation was to assess in vivo the ability of complementary angiotensin II (II Ang) peptides to block the biological effects of angiotensin II (Ang II). Increasing concentrations of rat or human II Ang were preincubated with Ang II for 2 hours, and this solution was then infused intra-arterially into the superior mesenteric artery. Human, but not rat, II Ang dose-dependently inhibited Ang II-induced mesenteric vasoconstriction. The in vivo inhibitory potencies of human II Ang and [Sar1,Ile8]Ang II, with respect to inhibition of the pressor response to Ang II, were compared by infusing intravenously increasing doses of each blocker and determining their effects on a fixed intravenous dose of Ang II. Although human II Ang could abolish the pressor response to Ang II, [Sar1,Ile8]Ang II was approximately 100 times more potent in this regard. A fixed dose of human II Ang (150 micrograms/min i.v.) inhibited the effects of increasing doses of Ang II on mesenteric vascular resistance, arterial blood pressure, and aldosterone secretion. The 1H nuclear magnetic resonance spectra of human II Ang and Ang II were determined both separately and when combined in the same cuvette. The spectrum obtained by overlaying the separate spectra for these two peptides was the same as the spectrum obtained from the mixture of these two peptides in the same cuvette.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗
PubMed1986

Regulation of pp60c-src synthesis by inducible RNA complementary to c-src mRNA in polyomavirus-transformed rat cells.

To determine the potential role of pp60c-src in polyomavirus-transformed cells, we constructed a recombinant plasmid with the mouse metallothionein-I promoter upstream of a src gene in an anti-sense orientation. We cotransfected this plasmid into middle tumor antigen-transformed FR3T3 cells with a plasmid containing the neomycin resistance gene, and G418 resistant colonies were selected. Analysis of these cells for pp60c-src expression revealed that 50 of the 200 cellular clones screened were found to have decreased levels of c-src expression when compared with the parental middle tumor antigen-transformed cells. Three independent clones which transcribed the expected 3.6-kilobase src complementary RNA and had levels of pp60c-src kinase activity comparable to that of normal FR3T3 cells were further analyzed. In the presence of Cd2+, these clones grew significantly slower in monolayer cultures than either the parental transformed cells (FR18-1) or FR18-1 cells transfected with the neomycin resistance gene alone. The morphology of these clones in the presence of Cd2+ was distinct from that of either the parental FR18-1 cells or normal FR3T3 cells. The clones expressing the complementary src RNA were found to form fewer colonies in soft agar, form fewer foci on monolayers of normal rat cells, and form tumors more slowly following injection into syngenic rats when compared with parental FR18-1 cells. The results of these studies suggest that the level of pp60c-src kinase activity affects the growth characteristics and transformation properties of polyoma virus-transformed rat cells.

Animals↗
PubMed1989

Monoclonal antibody against a peptide specified by [Met]-enkephalin complementary RNA recognizes the delta-class opioid receptor.

Monoclonal antibodies (MAB) were developed which recognize a peptide, His-Glu-Ala-Pro-Ile (HEAPI), encoded by the RNA complementary to the mRNA specifying [Met]-enkephalin. One such MAB (designated 6193) exhibited a high degree of reactivity to the peptide sequence. Other characteristics of 6193 MAB include: the ability to block opioid ligand binding in a radioreceptor assay; agonist activity similar to opioid peptides in suppressing cAMP production; and the recognition of a 58 kDa protein on the surface of the neuroblastoma x glioma cell line, NG108-15. These results are consistent with a reactivity of 6193 MAB with the delta-class opioid receptor.

Animals↗
PubMed1987

RNA complementary to alfalfa mosaic virus RNA 4 is not translated in vitro.

cDNA of the subgenomic messenger RNA (RNA 4) of alfalfa mosaic virus was ligated in both orientations into plasmids containing the bacteriophage SP 6 promoter. Upon transcription in vitro these plasmids yielded synthetic full-length complementary RNA and messenger-sense RNA. The complementary RNA contained an open reading frame of 417 nucleotides; the messenger-sense RNA encodes the virus coat protein. No translation products from the complementary RNA were detected in a wheat germ translation system, a reticulocyte lysate translation system, or Xenopus oocytes, whereas the messenger sense RNA was very active in these translation systems.

Autoradiography↗
PubMed1988

Purification of an angiotensin II binding protein by using antibodies to a peptide encoded by angiotensin II complementary RNA.

We have generated a monospecific antibody to a synthetic peptide encoded by an RNA complementary to the mRNA for angiotensin II (AII) and determined whether this antibody recognizes the AII receptor. We demonstrate that the antibody competes specifically with 125I-labeled AII for the same binding site on rat adrenal membranes. Furthermore, we show that this antibody inhibits the secretion of aldosterone from cultured rat adrenal cells, suggesting that the antibody recognizes the biologically relevant AII receptor. Finally, we demonstrate that antibody to the complementary peptide can be used to immunoaffinity-purify a protein of Mr 66,000 that specifically binds radiolabeled AII.

Adrenal Glands↗
PubMed1992

Expression of an outward-rectifying potassium channel from maize mRNA and complementary RNA in Xenopus oocytes.

Injection of Xenopus oocytes with poly(A)+ mRNA isolated from different plants (maize, cucumber, and squash) results in the appearance of a voltage- and time-dependent, potassium-selective, outward current that is similar to the outward-rectifying potassium current recorded in many higher plant cells. Maize shoots were found to be especially enriched in mRNA encoding such activity. A cDNA library of maize shoot mRNA was constructed in the vector lambda ZAPII and was used to synthesize RNA complementary to the cDNA (cRNA). Injection of the cRNA gave rise to an outward-rectifying potassium current with properties similar to the currents obtained by poly(A)+ mRNA injection. These results demonstrate that higher plant mRNA can be properly translated into a product that produces a voltage-regulated potassium channel in the plasma membrane of Xenopus oocytes. Thus, Xenopus oocytes can be used as a heterologous expression system for the functional identification and isolation of plant ion channel genes as well as for the study of structure-function relationship of plant ion channels.

Animals↗
PubMed1996

Detection of extended distribution of beta2-thyroid hormone receptor messenger ribonucleic acid (RNA) in adult rat brain using complementary RNA in situ hybridization histochemistry.

In situ hybridization histochemistry using a complementary RNA probe, directed at the entire length of the thyroid hormone receptor beta2 (TRbeta2)-specific region (-56 to 495 bp), was used to evaluate expression of TRbeta2 messenger RNA (mRNA) in coronal sections of adult rat brain. An extended distribution and intense expression of TRbeta2 mRNA was found in several regions of rat brain, including regions where TRbeta2 mRNA had not been previously identified using PCR or in situ hybridization histochemistry. These areas included hippocampus (dentate gyrus and C1, -2, and -3), cerebral cortex (predominantly layer 3), arcuate nucleus, median eminence, medial geniculate nucleus, tegmental bundle, medial and lateral lemniscus, Purkinje layer of the cerebellum, and several brain stem nuclei. In conclusion, we have developed a highly sensitive and specific method to demonstrate TRbeta2 mRNA expression in adult rat brain. The present findings are in agreement with immunoreactive TRbeta2 studies of rat brain and argue against the presence of an unidentified T3-binding protein to explain the previous discordant results of TRbeta2 mRNA and protein studies. In addition, the specificity of distribution of TR beta2 mRNA to certain brain nuclei, particularly those involved in hearing, implies a specific functional role of this receptor subtype and provides a physiological basis to understand the effects of hypothyroidism on brain development.

Animals↗
PubMed1986

Antibodies to the binding site of the receptor for luteinizing hormone-releasing hormone (LHRH): generation with a synthetic decapeptide encoded by an RNA complementary to LHRH mRNA.

A molecular recognition code has been hypothesized to exist in which ligands and their binding sites are encoded on complementary segments of genomic DNA. We have tested this hypothesis by generating a rabbit antibody to a synthetic decapeptide (complementary peptide) encoded by an RNA complementary to the mRNA for luteinizing hormone-releasing hormone (LHRH) and determining whether this antibody recognizes the LHRH receptor. When the antibody was used for immunoperoxidase staining of enzymatically dispersed rat anterior pituitary cells, only those that contained and secreted luteinizing hormone (i.e., the gonadotropes) were recognized. This staining could be abolished by preincubation with the complementary peptide or with an LHRH agonist, suggesting that the antibody is specific to the complementary peptide and is directed at the binding site of the receptor. Further evidence that the antibody recognizes the LHRH receptor was obtained in immunoblot experiments on solubilized receptors from pituitary glands. Immunoperoxidase staining with the antibody revealed two bands at 60 kDa and 51 kDa, which are values similar to those previously obtained for the LHRH receptor in photoaffinity-labeling experiments. The staining of these bands was inhibited by preincubation with the complementary peptide or an LHRH agonist. The antibody as well as the complementary peptide to LHRH also suppressed LHRH-stimulated luteinizing hormone release in a quantitative reverse hemolytic plaque assay, presumably by binding to the LHRH receptor and by binding LHRH, respectively. These findings suggest that the synthetic decapeptide whose sequence is specified by the complementary RNA to LHRH mRNA is sufficiently similar to an LHRH binding site that the peptide not only binds LHRH but was also recognized by the immune system as such a site. These findings provide strong support for the hypothesis that recognition molecules are encoded by complementary segments of genomic DNA.

Amino Acid Sequence↗
PubMed1993

An in vivo study of the replication origin in the influenza virus complementary RNA.

A new in vivo replication system for influenza virus was developed by using the clone 76 cell line, in which the viral RNA polymerase and nucleoprotein genes can be expressed in response to dexamethasone. The chimeric NS-chloramphenicol acetyltransferase (CAT) RNAs in the sense and antisense orientations positioned between the 5'- and 3'-terminal sequences of the influenza virus RNA segment 8 can be replicated [both genomic RNA (vRNA) and complementary RNA (cRNA) were transcribed] in the clone 76 cells treated with dexamethasone. These data indicate that three RNA polymerase proteins (PB1, PB2, and PA) and nucleoprotein are sufficient for replication of the influenza virus genome. Analysis of mutant cRNAs containing a base-substitution or a deletion in the 3'-conserved terminal 13 nucleotides revealed that important cis elements in the cRNA for vRNA synthesis reside at positions 2, 3, and 7 to 13 nucleotides from the 3'-end.

Animals↗
PubMed1999

The mitochondrial RNA ligase from Leishmania tarentolae can join RNA molecules bridged by a complementary RNA.

A biochemical characterization was performed with a partially purified RNA ligase from isolated mitochondria of Leishmania tarentolae. This ligase has a K(m) of 25 +/- 0.75 nM and a V(max) of 1.0 x 10(-4) +/- 2.4 x 10(-4) nmol/min when ligating a nicked double-stranded RNA substrate. Ligation was negatively affected by a gap between the donor and acceptor nucleotides. The catalytic efficiency of the circularization of a single-stranded substrate was 5-fold less than that of the ligation of a nicked substrate. These properties of the mitochondrial RNA ligase are consistent with an expected in vivo role in the process of uridine insertion/deletion RNA editing, in which the mRNA cleavage fragments are bridged by a cognate guide RNA.

Adenosine Triphosphate↗
PubMedDate not supplied

A novel immune system against bacteriophage infection using complementary RNA (micRNA).

The "operon' theory of gene regulation, in which protein repressor molecules bind to the operator site of a gene to prevent its transcription, is now well established. Recently, however, cases have been discovered in which gene expression is regulated by complementary RNA molecules that are able to bind to the transcripts of particular genes and consequently prevent their translation. For example, the synthesis of OmpF protein (a major outer membrane protein) in Escherichia coli is regulated by a short RNA complementary to a region of ompF RNA encompassing the "Shine-Dalgarno' sequence and the translation initiation codon. This RNA has been termed micRNA (messenger-RNA-interfering complementary RNA), and its discovery has prompted us to construct an artificial micRNA system designed to regulate gene expression in E. coli. A given target gene can be repressed by artificially producing an RNA (micRNA) complementary to the mRNA encoded by that gene. A micRNA system has also been used successfully in tissue-cultured mammalian cells. The use of artificial micRNAs to specifically regulate individual genes has great potential as a novel cellular immune system for blocking bacteriophage or virus infection. Here, we report that on induction of micRNAs directed against the coat protein and/or the replicase of the E. coli bacteriophage SP, phage proliferation was effectively prevented. We propose that the micRNA immune system provides an effective means of preventing viral infection as well as the expression of harmful genes in both prokaryotes and eukaryotes.

Base Sequence↗
PubMed1985

In vitro synthesis of full-length influenza virus complementary RNA.

Influenza virus-specific RNA has been synthesized in vitro, using cytoplasmic or microsomal fractions of influenza virus-infected MDCK cells. The RNA polymerase activity was stimulated 5-30 times by priming with ApG. About 20-30% of the product was polyadenylated. Most of the in vitro product was of positive polarity, as shown by hybridization to strand specific probes and by T1 fingerprinting of the poly(A)+ and poly(A)- RNA segments encoding haemagglutinin and nucleoprotein. The size of poly(A)- RNA segments, determined on sequencing gels, was indistinguishable from that of virion RNA, whereas poly(A)+ RNA segments contain poly(A) tails approximately 50 nucleotides long. The size of in vitro synthesized RNA segments was also determined by gel electrophoresis of S1-treated double-stranded RNAs, obtained by hybridization of poly(A)+ or poly(A)- RNA fractions with excess of unlabelled virion RNA. The results of these experiments indicate that poly(A)- RNA contains full-length complementary RNA. This conclusion is further substantiated by the presence of additional oligonucleotides in the T1 fingerprints of in vitro synthesized poly(A)- haemagglutinin or nucleoprotein RNA, selected by hybridization to cloned DNA probes corresponding to the 3' termini of the genes.

Animals↗