Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.21.64 (proteinase K)
 4,071 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The virion host shutoff (vhs) gene of herpes simplex virus encodes a virion polypeptide that induces degradation of host mRNAs at early times and rapid turnover of viral mRNAs throughout infection. To better investigate the vhs function, an in vitro mRNA degradation system was developed, consisting of cytoplasmic extracts from HeLa cells infected with wild-type herpes simplex virus type 1 or a mutant encoding a defective vhs polypeptide. Host and viral mRNAs were degraded rapidly in extracts from cells productively infected with wild-type herpes simplex virus type 1 but not in extracts from mock-infected cells or cells infected with the mutant vhs1. In contrast, 28S rRNA was stable in all three kinds of extract. Accelerated turnover of host mRNAs was also observed in extracts from cells infected with wild-type virus in the presence of dactinomycin, indicating that the activity was induced by a structural component of the infecting virions. The in vitro vhs activity was inactivated by heat or proteinase K digestion but was insensitive to brief treatment of the extracts with micrococcal nuclease. It was not inhibited by placental RNase inhibitor, it exhibited a strong dependence upon added Mg2+, it was active at concentrations of K+ up to 200 mM, and it did not require the components of an energy-generating system. In summary, the in vitro mRNA degradation system appears to accurately reproduce the vhs-mediated decay of host and viral mRNAs and should be useful for studies of the mechanism of vhs action.
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PMID:In vitro mRNA degradation system to study the virion host shutoff function of herpes simplex virus. 184 79

Messenger RNAs in eukaryotic cells exhibit a broad range of stabilities in vivo. Globin mRNA has a half life in excess of 50 h, but the half life of the c-myc oncogene mRNA is less than 20 min. Regulation of gene expression may be accomplished by a variety of mechanisms, including altering mRNA stability. We have examined the nuclear and cytoplasmic fractions of cells for factors affecting the metabolism of mRNA. Here we report that a HeLa whole-cell extract contains a factor that protects beta-globin mRNA from attack by RNases in a mouse erythroleukemia cell cytoplasmic extract. The factor is non-dialysable, inactivated by proteinase K and heat treatment, and resistant to RNase and DNase digestion. The HeLa cell factor resembles placental RNase inhibitor in that the mRNA-protecting activity is effective against RNase A and that treatment of the extract with N-ethylmaleimide completely destroys the protective activity. However, purified placental RNase inhibitor was unable to inhibit the RNase activity in the MELC cytoplasmic extract. These results suggest that the HeLa cell extract contains an RNase inhibitor (or inhibitors) with an activity or specificity that is distinct from that of placental RNase inhibitor.
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PMID:Cellular factor affecting the stability of beta-globin mRNA. 316 61

During or immediately after transcription of chromatin the high molecular weight pre-mRNA is complexes with proteins and low molecular weight RNA (lmwRNA). In the presence of a cytosolic RNase inhibitor pre-mRNA-protein complexes, designated as polyparticles, can be isolated from rat liver nuclei. The polyparticles are characterized by a maximum of their sedimentation coefficient of around 90 S, a protein to RNA ratio of 4.1, and a density in CsCl of 1.4 g/cm3. A set of 6--10 basic proteins of molecular weights between 30 and 45 kd as well as a multitude of polypeptides of higher molecular weights is associated with the rapidly labeled, polydisperse, high molecular weight RNA and several lmwRNA species. In order to study the structure of these very complex nuclear RNP complexes, the polyparticles were incubated at various concentrations of sodium chloride, urea or proteinases of different specificities (trypsin, chymotrypsin, proteinase K), recentrifuged through a sucrose layer and analyzed with respect to their sedimentation behavior, their protein to RNA ratios and their protein- and RNA components. Rhe results of these experiments led us to the proposal of a structural model which is presented here.
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PMID:The structure of ribonucleoprotein particles from rat liver nuclei. 701 68