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)

Heterogeneity of 5' untranslated region (5'UTR) sequences is a common feature of growth hormone receptor/binding protein (GHR/BP) mRNA from a number of species. Two major 5'UTR sequences (designated L1 and L2 in the mouse) have been cloned from rodents, ruminants and primates, and are known to correspond to transcripts generated from independently regulated promoters. A variable number of other 5'UTRs with diverse sequences have been cloned from rat, human and bovine tissues. To characterize alternative 5'UTR usage in mouse GHR/BP mRNA, we carried out 5' rapid amplification of cDNA ends using RNA from non-pregnant mouse liver and adipose tissue. Three novel 5'UTR sequences were obtained. Sequencing of genomic DNA revealed that exons corresponding to these three sequences are clustered within 1 kb downstream of the exon encoding 5'UTR L2, and the associated L2 promoter. The novel 5'UTRs are present at very low levels relative to the total pool of GHR/BP mRNA in liver, fat, kidney, and mammary gland as determined by ribonuclease protection assays. On the basis of these data, we propose that these 5'UTR sequences may result from the use of cryptic transcription start sites and splice donor sites under the influence of the adjacent L2 promoter/enhancer region.
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PMID:Alternative 5'-untranslated regions of mouse GH receptor/binding protein messenger RNA are derived from sequences adjacent to the major L2 promoter. 1101 62

Effects of d(CAG)(n).d(CTG)(n) repeats on expression of a reporter gene in human cell culture were studied using transient transfection, RNase protection and coupled transcription/translation assays. Cloning these repeats into the reporter 3'-UTR did not affect gene functioning. In contrast, placing the repeats in the reporter 5'-UTR led to strong inhibition of expression. This inhibition depended on the repeat orientation, being prominent only when the (CTG)(n) tracts were in the sense strand for transcription. Further, the strength of inhibition increased exponentially with an increase in repeat length. Our data indicate that expanded (CTG)(n) repeats prevent efficient translation of the reporter mRNA both in vitro and in vivo. We suggest that formation of stable hairpins by (CUG)(n) runs of increasing length in the 5'-UTR of a mRNA progressively inhibits the scanning step of translation initiation. This points to a novel mechanism of regulating gene expression by expandable d(CTG)(n) repeats.
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PMID:Expansion of the (CTG)(n) repeat in the 5'-UTR of a reporter gene impedes translation. 1102 74

We have cloned the mouse GDNF cDNA and genomic DNA to study the molecular mechanism of gene expression. Primer extension and RT-PCR analyses indicated that the mouse gene contains 1086 bp of 5'-untranslated region (5'-UTR) [Gene 203 (1997) 149]. In this report, we identified the core promoter region of mouse GDNF and examined the role of the 5'-UTR in gene expression. Promoter deletion analyses indicated that the proximal region (-81 to +28), which includes a TATA-box, is necessary for high-level expression of GDNF. Using reporter constructs encoding luciferase or fusion gene of GDNF to enhanced green fluorescent protein that were transiently transfected to mouse astroglial cell-line TGA-3 cells and rat glioma C6 cells, we investigated effects of the 5'-UTR on promoter activity. Luciferase reporter assay indicated that a region downstream of the transcription initiation site may include a positive regulatory element, while two more distal regions appear to contain negative regulatory elements, which was correlated to the mRNA level based on RNase protection assay. Both negative regulatory elements attenuated promoter activity in a position-dependent manner. Nuclear proteins from C6 glioma cells were shown to interact with several regions (+65/+105, +233/+265, and +554/+582) including each of the regulatory elements, suggesting that regulation of GDNF expression by the 5'-UTR occurred mainly at the transcriptional level.
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PMID:Promoter analysis and characteristics of the 5'-untranslated region of the mouse glial cell line-derived neurotrophic factor gene. 1114 11

Posttranscriptional regulation plays an important role in alpha-myosin heavy chain (alpha-MyHC) protein synthesis in cardiac muscle cells. In the present study, we test the effects of calcium and mRNA mislocalization on alpha-MyHC translation in order to determine the mechanism(s) contributing to translational block via the 3' untranslated region (3'UTR). Neonatal rat cardiac myocytes were treated for 6 h with L-isoproterenol (10 microM) to enhance beating, with 10 microM verapamil to block beating and mislocalize mRNA, or with 3 microM colchicine to enhance beating but mislocalize mRNA by depolymerization of the microtubules. In order to determine whether translation is regulated by the 3'UTR, either a control (SV40 3'UTR) or the experimental (alpha-MyHC 3'UTR) was placed after a luciferase reporter gene and transfected into the myocytes. The amount of luciferase protein only decreased significantly in verapamil arrested cells transfected with the alpha-MyHC 3'UTR construct (P < 0.01). To control for the possibility that pharmacological treatments might affect transcription or message stability, we analyzed neomycin and luciferase mRNA levels transcribed from the same transfected plasmid. No significant changes were found with an RNase protection assay. These results suggest that calcium but not mRNA localization regulates protein synthesis and further, this is mediated by the 3' untranslated region of alpha-MyHC.
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PMID:Translation is regulated via the 3' untranslated region of alpha-myosin heavy chain mRNA by calcium but not by its localization. 1120 37

Analysis of the Haloarcula marismortui large ribosomal subunit has revealed a common RNA structure that we call the kink-turn, or K-turn. The six K-turns in H.marismortui 23S rRNA superimpose with an r.m.s.d. of 1.7 A. There are two K-turns in the structure of Thermus thermophilus 16S rRNA, and the structures of U4 snRNA and L30e mRNA fragments form K-turns. The structure has a kink in the phosphodiester backbone that causes a sharp turn in the RNA helix. Its asymmetric internal loop is flanked by C-G base pairs on one side and sheared G-A base pairs on the other, with an A-minor interaction between these two helical stems. A derived consensus secondary structure for the K-turn includes 10 consensus nucleotides out of 15, and predicts its presence in the 5'-UTR of L10 mRNA, helix 78 in Escherichia coli 23S rRNA and human RNase MRP. Five K-turns in 23S rRNA interact with nine proteins. While the observed K-turns interact with proteins of unrelated structures in different ways, they interact with L7Ae and two homologous proteins in the same way.
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PMID:The kink-turn: a new RNA secondary structure motif. 1148 24

The 5' untranslated region (5'UTR) of the psbA mRNA (psbA encodes the PSII reaction center protein, D1) is a key site for RNA-protein interactions in the post-transcriptional regulation of gene expression. In this study, we mapped the major psbA mRNA 5'-terminus at -77 nt, and two minor termini clusters centered at -48 and -64 nt, upstream from the psbA translational start codon of Arabidopsis thaliana. RNA mobility shift, RNase protection and UV-crosslinking assays were used to characterize the interaction of chloroplast proteins with the RNA 5'UTR. RNA-protein interactions depended upon a thermolabile secondary structure and specific sequences in a 35 nt region of the 5'UTR, which were 80% conserved with the psbA 5'UTRs from five other plants. Major and minor proteins of 43- and 30-kDa, respectively, were detected by UV-crosslinking to RNA. Oxidizing conditions abolished the association of the proteins with the 5'UTR, while RNA-binding activity was recovered upon incubation with a reductant. Based on these findings, we hypothesize that post-transcriptional regulation of psbA gene expression in chloroplasts of vascular plants involves redox-dependent interactions between specific sequences in the 5'UTR and 43- and 30-kDa RNA-binding proteins.
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PMID:RNA binding-proteins interact specifically with the Arabidopsis chloroplast psbA mRNA 5' untranslated region in a redox-dependent manner. 1167 22

A range of isolates of Pea seed-borne mosaic virus (PSbMV) was compared in the segments of the genome representing the partial NIb/CP/UTR and the partial P1-Pro/HC-Pro coding regions. Nucleotide and amino acid sequences, and a phylogenetic analysis of the CP region, divided isolates with available sequence information into two groups, one representing pathotype 4, the other pathotype 1. The pathotype 1 group showed greater diversity than the pathotype 4 group. A comparison of 14 isolates, S6 (a pathotype 4 isolate), US (a pathotype 1 isolate) and 12 isolates from Pakistan, by ribonuclease protection assay (RPA) using cRNA transcripts of the cloned partial NIb/CP/UTR regions of the S6, US and Pakistani isolate PK9 placed them into three distinct phylogenetic groups. RPA with a partial P1-Pro/HC-Pro cRNA probe identified a greater level of variation which was too high to be used for generating an overall phylogeny. Thus, RPA identified greater molecular diversity in PSbMV than described hitherto. We conclude that, in addition to the pathotypes 1 and 4 typified by US and S6 respectively, isolates of PSbMV from Pakistan include previously unrecognised molecular variants, and this accords with our previous recognition of new pathotypes from Pakistan.
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PMID:Genomic heterogeneity in Pea seed-borne mosaic virus isolates from Pakistan, the centre of diversity of the host species, Pisum sativum. 1172 10

Neuroserpin is an axonally secreted serine protease inhibitor expressed in the nervous system that protects neurons from ischemia-induced apoptosis. Mutant neuroserpin forms have been found polymerized in inclusion bodies in a familial autosomal encephalopathy causing dementia, or associated with epilepsy. Regulation of neuroserpin expression is mostly unknown. Here we demonstrate that neuroserpin mRNA and the RNA-binding protein HuD are co-expressed in the rat central nervous system, and that HuD binds neuroserpin mRNA in vitro with high affinity. Gel-shift, supershift and T1 RNase assays revealed three HuD-binding sequences in the 3'-untranslated region (3'-UTR) of neuroserpin mRNA. They are AU-rich and 20, 51 and 19 nt in length. HuD binding to neuroserpin mRNA was also demonstrated in extracts of PC12 pheochromocytoma cells. Additionally, ectopic expression of increasing amounts of HuD in these cells results in the accumulation of neuroserpin 3'-UTR mRNA. Furthermore, stably transfected PC12 cells over-expressing HuD contain increased levels of both neuroserpin mRNAs (3.0 and 1.6 kb) and protein. Our results indicate that HuD stabilizes neuroserpin mRNA by binding to specific AU-rich sequences in its 3'-UTR, which prolongs the mRNA lifetime and increases protein level.
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PMID:HuD binds to three AU-rich sequences in the 3'-UTR of neuroserpin mRNA and promotes the accumulation of neuroserpin mRNA and protein. 1200 Aug 40

Mitochondrial NAD(+)-dependent succinic semialdehyde dehydrogenase (ALDH5A1, SSADH) represents the last enzyme in the GABA catabolism and irreversibly oxidizes SSA to succinate. In human, SSADH deficiency results in 4-hydroxybutyric aciduria, an autosomal recessive disorder due to an accumulation of GABA and 4-hydroxybutyric acid in the CNS. We already identified SSADH gene on human chromosome 6p22 and characterized the coding region. Furthermore, we described the first two mutations causing the disease. We report here the complete cDNA and genomic structure of the gene. A single transcription start site was identified by RNase protection 122 bp upstream of the ATG. EST database search and reporter gene constructs of the 3(') genomic region showed that the two major SSADH mRNA isoforms are due to alternative polyadenylation sites. The two mRNAs of 1827 and 5225 nt were analyzed for differential stability and translation efficiency. The analysis of mRNA turnover showed that both SSADH transcripts are equally stable. Similarly, a measurement of polysomal association capability of the two GFP-SSADH reporter mRNAs (containing the 3' UTR regions of the two SSADH mRNAs) did not reveal any difference. However, we cannot exclude the fact that differential properties could be restricted to particular physiological conditions and/or specific tissues. We have also identified an alternatively spliced small exon, which may lead to a novel isoform of the enzyme. Furthermore, we report here on naturally occurring missense variants, which may significantly contribute to inter-individual variation of SSADH activity, possibly influencing GABA and GHB endogenous levels.
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PMID:Structure of human succinic semialdehyde dehydrogenase gene: identification of promoter region and alternatively processed isoforms. 1220 42

Stabilization and correct localization of mRNA are important features of renin synthesis. To elucidate the molecular basis of cAMP-mediated posttranscriptional control via mRNA stabilization, we analyzed the interaction of human preprorenin (hREN) mRNA 3'-untranslated region (3'-UTR) with proteins of renin synthesizing Calu-6 cells and investigated their functional impact on messenger integrity. To identify hREN mRNA binding proteins, electrophoretic mobility shift assays, UV cross-linking and RNA-affinity chromatography with subsequent matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were performed. The following six proteins were unambiguously identified as hREN mRNA 3'-UTR binding proteins: hnRNP E1 (synonyms alpha-CP or PCBP), hnRNP K, dynamin, nucleolin, YB-1, and MINT-homologous protein. All proteins contain various RNA binding motifs, and most have been described in the context of mRNA binding and mRNA stabilization. Four proteins for which antibodies were available were verified by immunological techniques (dynamin, nucleolin, hnRNP E1, and YB-1). Forskolin, an activator of cAMP synthesis, considerably stimulates renin synthesis via inhibition of REN mRNA decay. Functionally, this cAMP-based mRNA stabilization is accompanied by a 3- to 6-fold upregulation of REN mRNA binding proteins. RNase degradation assays confirm that 3'-UTR binding proteins are able to protect and stabilize REN mRNA in vitro.
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PMID:Posttranscriptional control of renin synthesis: identification of proteins interacting with renin mRNA 3'-untranslated region. 1260 Aug 97


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