Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.27.1 (RNase)
 16,360 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The structure of full-length rat prodynorphin cDNA and the corresponding gene has been determined. The 2400 base rat prodynorphin mRNA is encoded by four exons. Exons 1 and 2 encode the majority of the 5'-untranslated sequence, while exons 3 and 4 contain the translated region; the entire 3' -untranslated region is contained on exon 4 as well. RNase protection studies, in which a genomic DNA fragment was used to generate a cRNA hybridization probe, have determined the major transcriptional initiation site for both brain and testicular prodynorphin mRNA. Transient expression of transfected fusion genes containing the 5'-flanking DNA of the rat prodynorphin gene linked to the structural sequence of a reporter gene has been used to identify specific genomic DNA fragments from the prodynorphin gene locus which are capable of acting as transcriptional promoters. Multiple regions of genomic DNA appear to have transcriptional promoter activity when introduced into various eukaryotic cell lines.
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PMID:Characterization of the rat prodynorphin gene. 262 41

Prodynorphin gene transcripts have been characterized in human tissues with cRNA probes covering parts of the non-coding exon 1 and the main coding exon 4 using Northern blot and RNase protection experiments. A 2.8-kb signal was observed in human brain RNA with both the exon 1 and exon 4 probes. An RNase protection assay with the exon 1 probe, performed to map the 5' end of this transcript, produced protected fragments in the range of 0.11 to 0.15 kb indicating that exon 1 is 1.2 kb shorter than previously proposed. 5'-RACE-PCR and sequencing of amplified cDNA fragments confirmed this assignment. In adrenal gland, testis and the human small cell lung carcinoma cell line, U1690, several prodynorphin-like mRNAs structurally different from the brain prodynorphin mRNA species were observed.
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PMID:Characterization of human prodynorphin gene transcripts. 748 56

The solution hybridization RNase protection assay may be considered a suitable method for both qualitative and quantitative analysis of mRNAs. In the present study we described an application of solution hybridization RNase protection assay to the quantitative analysis of prodynorphin mRNA, which encodes for the synthesis of prodynorphin, a common precursor for a number of opioid peptides. In the myocardial cell, stimulation of the K opioid is involved in the modulation of cytosolic calcium and pH homeostasis. In the present study, we found that prodynorphin mRNA, which encodes for the synthesis of a common precursor of opioid peptides interacting with K sites, is synthesized both in atrial and in ventricular tissue of the rat heart. In adult cultured rat ventricular cardiomyocytes, the level of prodynorphin mRNA did not differ from that detected in the original ventricular tissue. This finding indicates that the myocardial cell is an important source for prodynorphin gene expression and has the potential for an intrinsic synthesis of dynorphin-related peptides.
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PMID:Evaluation of opioid peptide gene expression by solution hybridization RNase protection. 789 79

The expression of the prodynorphin gene was investigated in adult cultured rat ventricular cardiac myocytes by using a sensitive solution hybridization RNase protection assay for the quantitative analysis of prodynorphin mRNA. Myocyte culture in high KCl resulted, after 4 h, in a marked increase in cellular prodynorphin mRNA, while a KCl treatment for 6, 12, or 24 h progressively down-regulated the levels of prodynorphin mRNA below the control value. Immunoreactive dynorphin B, a biologically active end product of the precursor, was found to be present in the culture medium in significantly higher amounts than in the cardiac myocytes. The levels of this biologically active K opioid receptor agonist significantly increased after 4 h of KCl treatment and were markedly reduced following a 24-h exposure of the cardiac myocytes to KCl. These KCl-induced effects were all abolished by cell incubation in the presence of the calcium channel blocker verapamil. In single cardiac myocytes, acute stimulation of K opioid receptors with dynorphin B or with the selective agonist U-50,488H increased the level of cytosolic calcium. This effect was abolished by the specific K opioid receptor antagonist (Mr-1452) and was not affected by the removal of calcium from the bathing medium. These results suggest that an opioid gene may influence the myocardial function in an autocrine or paracrine fashion.
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PMID:Dynorphin gene expression and release in the myocardial cell. 790 74

Preprodynorphin and preproenkephalin are protein precursors from which are derived two classes of opioid neurotransmitter peptides. Dynorphin A((1-17)) is produced by proteolytic processing of prodynorphin, and processing of proenkephalin yields the enkephalin peptides. We report here on the isolation and sequencing of multiple clones for these two mRNAs from a cDNA library. Two cDNA clones of preprodynorphin contained the full-length sequence (2.35 kb) with the primary structure predicted from the guinea pig gene sequence. In contrast, one clone encoded the full-length sequence but also an additional 192 nt at the 5' end. This sequence has high homology to the 5' flanking region of the human preprodynorphin gene, and RNase protection assays demonstrated that in addition to a primary initiation site, transcription of this mRNA is initiated at several sites 160-190 nt 5' with respect to the primary site. This difference may alter translational efficiency or mRNA stability. The sequence of preproenkephalin cDNA clones confirmed the structure predicted from the gene sequence. One clone, however, contained sequences encoded by exons 2 and 3, and initiated within the first intron (intron A) of the gene. We used RNase protection mapping to assess the abundance in the brain and pituitary of preproenkephalin transcripts that initiate within intron A. These studies confirmed that the primary transcription start site is 28 nucleotides downstream from the TATAA site, and that intron A sequences are not present in significant amounts in these tissues.
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PMID:Primary structure of guinea pig preprodynorphin and preproenkephalin mRNAs: multiple transcription initiation sites for preprodynorphin. 1513 Jul

We show that temperature is an important parameter for the sensitivity of saturation transfer difference (STD) spectroscopy. A decreased intensity of STD signals is observed for lactose binding to growth-regulatory galectin7 (p53-induced gene 1), as well as for nucleotide binding to annexin A6, when the temperature is increased from 281 to 298-310 K. Opposite temperature effects on STD intensity are observed for S-peptide binding to S-protein to reconstitute RNase S. However, the STD signals for tryptophan binding to downstream regulatory element antagonist modulator of the human prodynorphin gene (DREAM)are relatively unaffected between 281 and 298 K. The known kinetics of the binding of ATP by the uncoupling protein from brown adipose tissue mitochondria (UCP1) predicted an observable STD at 310 K, but rapid sample degradation limits the experiments to much lower temperatures. Temperature strongly influences the kinetics and affinity constant of various types of complex formation and in so doing influences the observed STD effects. Therefore, temperature can be exploited to facilitate the optimization of STD-based applications, and at the same time minimize the number of test samples. STD-based screening protocols to detect new target-specific compounds may yield a larger number of potential ligands if screened at various temperatures.
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PMID:Temperature dependence of ligand-protein complex formation as reflected by saturation transfer difference NMR experiments. 1763 17