Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.13.1 (exoribonuclease)
 732 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The Saccharomyces cerevisiae OLE1 gene encodes the Delta-9 fatty acid desaturase, a highly regulated integral membrane enzyme involved in the formation of unsaturated fatty acids from saturated acyl-coenzyme A precursors. The mRNA levels of the OLE1 gene are regulated by at least two independent control systems that respond to nutrient fatty acids. One involves the unsaturated fatty acid repression of OLE1 transcription; the second, described in this report, involves unsaturated fatty acid-responsive changes in the half-life of the OLE1 mRNA. Measurements of OLE1 mRNA half-life indicate that it is a moderately stable species (t1/2 = 10 +/- 1.5 min) in cells grown in medium without exogenous fatty acids. Its half-life is drastically reduced (t1/2 < 2.5 min), in a time-dependent manner, following the addition of unsaturated fatty acids to the growth medium. Saturated fatty acids that have previously been shown to increase activation of OLE1 transcription do not regulate its mRNA stability. Inhibition of translation, by the addition of cycloheximide, slows the nucleolytic degradation of the OLE1 mRNA and blocks the unsaturated fatty acid-triggered reduction in its half-life. This suggests an intimate link between the two processes of mRNA decay and protein synthesis. A chimeric mRNA, produced by replacing the upstream activation and fatty acid-regulated regions of the OLE1 promoter with the GAL1 promoter sequences is destabilized by exogenous unsaturated fatty acids. A similar chimera under GAL1 control that replaces the OLE1 mRNA 5'-untranslated region with GAL1 sequences is not regulated by unsaturated fatty acids. These results suggest that the 5'-untranslated region of the OLE1 mRNA contains sequence elements required for fatty acid-triggered destabilization. Disruption of the XRN1 gene, which encodes a 5' --> 3'-exoribonuclease, results in an approximate 4-fold increase in OLE1 mRNA half-life in the absence of fatty acids. Its half-life is reduced when those cells are exposed to unsaturated fatty acids, indicating that the 5'-exoribonuclease encoded by the XRN1 gene is required for the rapid degradation of the OLE1 transcript but is not required for fatty acid-induced destabilization.
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PMID:Fatty acid-responsive control of mRNA stability. Unsaturated fatty acid-induced degradation of the Saccharomyces OLE1 transcript. 882 9

RNase II is a single-stranded-specific 3'-exoribonuclease that degrades RNA generating 5'-mononucleotides. This enzyme is the prototype of an ubiquitous family of enzymes that are crucial in RNA metabolism and share a similar domain organization. By sequence prediction, three different domains have been assigned to the Escherichia coli RNase II: two RNA-binding domains at each end of the protein (CSD and S1), and a central RNB catalytic domain. In this work we have performed a functional characterization of these domains in order to address their role in the activity of RNase II. We have constructed a large set of RNase II truncated proteins and compared them to the wild-type regarding their exoribonucleolytic activity and RNA-binding ability. The dissociation constants were determined using different single- or double-stranded substrates. The results obtained revealed that S1 is the most important domain in the establishment of stable RNA-protein complexes, and its elimination results in a drastic reduction on RNA-binding ability. In addition, we also demonstrate that the N-terminal CSD plays a very specific role in RNase II, preventing a tight binding of the enzyme to single-stranded poly(A) chains. Moreover, the biochemical results obtained with RNB mutant that lacks both putative RNA-binding domains, revealed the presence of an additional region involved in RNA binding. Such region, was identified by sequence analysis and secondary structure prediction as a third putative RNA-binding domain located at the N-terminal part of RNB catalytic domain.
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PMID:Characterization of the functional domains of Escherichia coli RNase II. 1680 66

The major cytoplasmic 5' to 3'-exoribonuclease activity is carried out by the Xrn1 protein in eukaryotic cells. A number of different approaches can be used to study multifunctional Xrn1 protein activity in vitro. In this chapter, we concentrate on methods used in our laboratory to analyze Xrn1 5' to 3'-exoribonuclease activity. Some of these techniques may also be suitable for detecting 3' to 5'-exoribonuclease or endoribonuclease activity. For these reasons, these assays can be used to isolate new proteins with ribonuclease activity and, when performed in combination with in vivo experiments, will contribute to a new level of understanding of the function of these factors.
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PMID:In vitro assays of 5' to 3'-exoribonuclease activity. 1911 Nov 76

RNase II is the prototype of a ubiquitous family of enzymes that are crucial for RNA metabolism. In Escherichia coli this protein is a single-stranded-specific 3'-exoribonuclease with a modular organization of four functional domains. In eukaryotes, the RNase II homologue Rrp44 (also known as Dis3) is the catalytic subunit of the exosome, an exoribonuclease complex essential for RNA processing and decay. In this work we have performed a functional characterization of several highly conserved residues located in the RNase II catalytic domain to address their precise role in the RNase II activity. We have constructed a number of RNase II mutants and compared their activity and RNA binding to the wild type using different single- or double-stranded substrates. The results presented in this study substantially improve the RNase II model for RNA degradation. We have identified the residues that are responsible for the discrimination of cleavage of RNA versus DNA. We also show that the Arg-500 residue present in the RNase II active site is crucial for activity but not for RNA binding. The most prominent finding presented is the extraordinary catalysis observed in the E542A mutant that turns RNase II into a "super-enzyme."
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PMID:Determination of key residues for catalysis and RNA cleavage specificity: one mutation turns RNase II into a "SUPER-ENZYME". 1945 82

Deadenylation of eukaryotic mRNA is a mechanism critical for mRNA function by influencing mRNA turnover and efficiency of protein synthesis. Here, we review poly(A)-specific ribonuclease (PARN), which is one of the biochemically best characterized deadenylases. PARN is unique among the currently known eukaryotic poly(A) degrading nucleases, being the only deadenylase that has the capacity to directly interact during poly(A) hydrolysis with both the m(7)G-cap structure and the poly(A) tail of the mRNA. In short, PARN is a divalent metal-ion dependent poly(A)-specific, processive and cap-interacting 3'-5' exoribonuclease that efficiently degrades poly(A) tails of eukaryotic mRNAs. We discuss in detail the mechanisms of its substrate recognition, catalysis, allostery and processive mode of action. On the basis of biochemical and structural evidence, we present and discuss a working model for PARN action. Models of regulation of PARN activity by trans-acting factors are discussed as well as the physiological relevance of PARN.
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PMID:Poly(A)-specific ribonuclease (PARN): an allosterically regulated, processive and mRNA cap-interacting deadenylase. 2349 18

Regulated mRNA decay plays a vital role in determining both the level and quality of cellular gene expression. Viral RNAs must successfully evade this host RNA decay machinery to establish a productive infection. One way for RNA viruses to accomplish this is to target the cellular exoribonuclease XRN1, because this enzyme is accessible in the cytoplasm and plays a major role in mRNA decay. Members of the Flaviviridae use RNA structures in their 5'- or 3'-untranslated regions to stall and repress XRN1, effectively stabilizing viral RNAs while also causing significant dysregulation of host cell mRNA stability. Here, we use a series of biochemical assays to demonstrate that the 3'-terminal portion of the nucleocapsid (N) mRNA of Rift Valley fever virus, a phlebovirus of the Bunyaviridae family, also can effectively stall and repress XRN1. The region responsible for impeding XRN1 includes a G-rich portion that likely forms a G-quadruplex structure. The 3'-terminal portions of ambisense-derived transcripts of multiple arenaviruses also stalled XRN1. Therefore, we conclude that RNAs from two additional families of mammalian RNA viruses stall and repress XRN1. This observation. emphasizes the importance and commonality of this viral strategy to interfere with the 5'-to-3'-exoribonuclease component of the cytoplasmic RNA decay machinery.
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PMID:Identification of phlebovirus and arenavirus RNA sequences that stall and repress the exoribonuclease XRN1. 2911 86