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Results for “RNA, Small Cytoplasmic”

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At least 19 recordsLinked to original sources

PubMed2025

EV-D68 cleaves LARP1 and PABPC1 by 3Cpro to redirect host mRNA translation machinery toward its genomic RNA.

Enterovirus D68 (EV-D68) is an emerging pathogen associated with severe respiratory diseases and neurological complications, such as acute flaccid myelitis. EV-D68 has developed sophisticated mechanisms to hijack host translation machinery, facilitating its replication and impairing host mRNA translation. In this study, we demonstrate that EV-D68 cleaves La-related protein 1 (LARP1) and poly(A)-binding protein cytoplasmic 1 (PABPC1) through its proteases 3Cpro and 2Apro. Our results indicate that overexpressing LARP1 and PABPC1 significantly inhibits EV-D68 replication and reduces the virus-mediated suppression of host translation. While both LARP1 and PABPC1 regulate translation, they exert antiviral effects through distinct mechanisms. We found that LARP1 interacts with the 5'UTR of EV-D68 RNA through its LAM domain, and this interaction is crucial for its antiviral function. LARP1 translation modulation is also influenced by the mTOR and CDK1 signaling pathways. Viral infection inhibits mTOR and CDK1 phosphorylation, which enhances LARP1's binding to viral RNA and inhibits viral translation. To counteract this inhibition, EV-D68 cleaves LARP1 through 3Cpro, thereby promoting efficient viral translation. We also investigated other enteroviruses, such as EV-A71 and CV-A16, which similarly target LARP1 and PABPC1, indicating a conserved mechanism across enteroviruses. Our findings offer new insights into how EV-D68 manipulates host translation and highlight the potential of targeting LARP1 and PABPC1 for antiviral interventions.

Humans↗

PubMed1975

Polyadenylic acid on poliovirus RNA. III. In vitro addition of polyadenylic acid to poliovirus RNAs.

A crude RNA polymerase preparation was made from HeLa cells infected for 3 h with poliovirus. All virus-specific RNA species labeled in vitro (35S RNA, replicative intermediate RNA [RI], and double-stranded RNA [dsRNA]) would bind to poly(U) filters and contained RNase-resistant stretches of poly(A) which could be analyzed by electrophoresis in polyacrylamide gels. After incubation for 45 min with [3-H]ATP in the presence of the other three nucleoside triphosphates, the labeled poly(A) on the RI and dsRNA migrated on gels as relatively homogenous peaks approximately 200 nucleotides in length. In contrast, the poly(A) from the 35S RNA had a heterogeneous size distribution ranging from 50 to 250 nucleotides. In the absence of UTP, CTP, and GTP, the size of the newly labeled poly(A) on the dsRNA and RI RNA was the same as it was in the presence of all four nucleoside triphosphates. However the poly(A) on the 35S RNA lacked the larger sequences seen when the other three nucleoside triphosphates were present. When [3-H]ATP was used as the label in infected and uninfected extracts, heterogeneous single-stranded RNA sedimenting at less than 28S was also labeled. This heterogeneous RNA probably represents HeLa cytoplasmic RNA to which small lengths of poly(A) (approximately 15 nucleotides) had been added. These results indicate that in the in vitro system poly(A) can be added to both newly synthesized and preexisting RNA molecules. Furthermore, an enzyme capable of terminal addition of poly(A) exists in both infected and uninfected extracts.

Adenine Nucleotides↗

PubMed1978

Histology and histochemistry of the normal superficial corneal epithelium of rabbit.

The superficial epithelial cells of normal cornea are in a process of gradual metabolic shutdown. Nuclear DNA may be depolymerized. Loss of DNA also occurs by extrusion in small quantities into the cytoplasm. There is a small quantity of RNA in these cells. The cytoplasm shows the presence of some vicglycols. The superficial corneal epithelial cells do not keratinize.

Animals↗

PubMed1978

Preparative density gradient centrifugation of RNA and DNA in cesium sulfate-urea mixture.

A new procedure is described for a large scale separation and purification of unfixed DNA and RNA from a mixture of partially extracted nucleic acids and lysates of subcellular fractions by centrifugation to equilibrium in cesium sulfate-urea mixture. Optimum conditions are described for the separation and quantative recovery of both RNA and DNA in a pure form. The procedure allows determination of peak buoyant densities of 4-5s RNA, 7-11s mRNA and total cytoplasmic RNA. The procedure also allows fractionation of small molecular weight classes of cytoplasmic RNAs from the 18s and 28s rRNAs.

Animals↗

PubMed1976

Maturation of ribosomes in yeast. II. Position of the low molecular weight rRNA species in the maturation process.

Yeast protoplasts were pulse labelled with [5-3H] uridine and the labelling kinetics of the low molecular weight rRNA species were determined in order to gain more insight into the position of those small rRNAs in the process of ribosome maturation. 7-S RNA, the immediate precursor of 5.8-S rRNA, is found to be present only in the nucleus, indicating that the conversion of 7-S to 5.8-S RNA is a nuclear event. 5.8-S rRNA is observed in the cytoplasm almost immediately after its formation. This as well as the presence of only a small amount of 5.8-S RNA in the nucleus, shows that the ribosomal precursor particles of the large ribosomal subunit are very rapidly transported into the cytoplasm once 5.8-S rRNA is formed. Most of the newly synthesized 5-S RNA is found in the nucleus. This nuclear 5-S rRNA is mainly present in the ribosomal precursor particles. However, a small pool of free 5-S rRNA is probably also present.

Cell Fractionation↗

PubMed1978

Characterization of two SV40 early mRNAs and evidence for a nuclear "prespliced" RNA species.

Using in vitro translation of sucrose-gradient fractionated cytoplasmic mRNA from SV40-infected cells, we have shown that a deletion in the region mapping between 0.54--0.59 reduced the size of mRNA for small-t but not the size of mRNA for large-T. Mutants with a deletion in this region were shown to produce in vivo either shortened small-t or no small-t, and normal large-T. Similarly, in vitro translation of poly(A)+cytoplasmic RNA from cells infected with these mutants gave the same results. On the other hand in vitro translation of poly(A)+nuclear RNA from the mutants which made no small-t produced a small-t derivative possibly synthesized from a prespliced RNA species. We have also shown that poly(A)+nuclear RNA from mutant dl 2122 produced two small-t related proteins: one of these (MW: 11K) probably represents the product of a "prespliced" RNA, the other (MW: 17K) which is also found in the cytoplasm represents the product of the mutant specific small-t mRNA.

Antigens, Neoplasm↗

PubMed1976

Quantitation of turnover and export to the cytoplasm of hnRNA transcribed in the Balbiani rings.

Some quantitative parameters of the intranuclear metabolism and export to the cytoplasm of Balbiani ring 1 and 2 RNA molecules in salivary gland cells of Chironomus tentans have been determined. Growing RNA chains in the Balbiani rings attain uniform labeling with RNA precursors after 20 min of incorporation. The specific activity of 75S RNA released from the Balbiani rings into the nuclear sap increases rapidly and reaches a maximum level between 90 and 180 min of labeling. After 20 min, labeled 75S RNA enters the cytoplasm and accumulates at a linear rate. However, only a small proportion of the RNA produced at the Balbiani ring loci can subsequently be recovered in the nuclear sap (14-17%) or cytoplasm (4-7%) as 75S RNA; presumably the remainder is degraded entirely. Experiments using inhibitors of elongation (actinomycin D) or initiation (5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole) revealed that no significant quanity of the 75S RNA transcribed can be chased into the cytoplasm. Both the kinetics of entry of labeled 75S RNA into the cytoplasm-that is, a constant rate of increase after a brief lag-and chase data are incompatible with a precursor-product relationship between the great majority of nuclear 75S RNA and cytoplasmic 75S RNA with messenger characteristics. The results are discussed in relation to the possibility that a post-transcriptional control mechanism is operating in these cells.

Animals↗

PubMed1979

Properties of a small transcribed poly A sequence in heterogeneous nuclear RNA of HeLa cells.

A class of heterogeneous nuclear RNA (hnRNA) molecules contain an internal transcribed poly A sequence of close to 25 uninterrupted AMP residues. HnRNA molecules containing this sequence are separable from those containing the large 3' terminal poly A sequence on the basis of their differential affinity for oligo dT cellulose. The fact that the transcribed small poly A and the 3' terminal poly A are not found in the same hnRNA molecules even though both are present in similar size classes and that the small poly A is absent from cytoplasmic messenger RNA (mRNA) has led us to propose a scheme for mRNA processing in which the 3' end of the small poly A in hnRNA becomes a priming size for the post-transcriptional addiction of the large poly A.

HeLa Cells↗

PubMed1975

Relationship between A-type and C-type particles in cells infected by Rous sarcoma virus.

Chicken cells infected with avian RNA tumor virus often contain small cytoplasmic A-type particles which commonly exist as clusters of 50--100 particles when viewed in thin sections. These particles were found more consistently in Rous sarcoma virus-infected than Rous-associated virus-infected cultures, but were generally present in only a small fraction of the total infected cells. The results of the survey of cells infected with various strains of leukosis-sarcoma viruses led to the hypothesis that the A particles develop in cells undergoing cytopathological degeneration. The hypothesis explains also the evanescent nature of the appearance of these particles in infected cells. The application of immunoelectron microscopic methods using monospecific antisera against viral internal proteins revealed that the A particles contain components immunologically related to the proteins of C-type virus.

Animals↗

PubMed1976

Small RNA species of the HeLa cell: metabolism and subcellular localization.

The small molecular weight RNAs of the HeLa cell have been located in specific subcellular fractions. SnA is located in the nucleolus and is partially bonded to nucleolar 28S RNA. SnD, the most abundant of the small nuclear RNAs, is partially released from the nucleus when the nuclear preparation is briefly warmed. SnF is released from the nuclei when chromatin is digested with the micrococcal nuclease and not when pancreatic DNAase is used. The remainder of the small nuclear species remain in the nucleus following the digestion of chromatin and are concluded to be elements of the "nuclear skeleton." SnK is found predominantly in the cytoplasm, but migrates quantitatively to the nuclear fraction in the presence of high levels of actinomycin D. ScL is totally cytoplasmic and is partially bound to cell membranes. It is the 7S RNA found in oncornavirus virions. All the small nuclear RNAs appear initially in the cytoplasmic fraction before fixation in the nucleus. Two short-lived cytoplasmic species behave kinetically as precursors to the stable nuclear RNAs.

Cell Fractionation↗

PubMed1977

Free and membrane-bound polyribosomes in BHK cells infected with Sindbis virus.

The data presented in the paper demonstrate that in BHK cells infected with Sindbis virus virtually all the 42S mRNA not in nucleocapsid is associated with free polyribosomes, whereas the 26S mRNA is distributed between free and membrane-bound polyribosomes. We suggest that the 26S RNA polyribosomes are bound to the membranes through the nascent chains of the B1 protein and that a large percentage of 26S RNA polyribosomes free in the cytoplasm may be due to the small amount of rough endoplasmic reticulum in BHK cells. In addition, we found that intracellular nucleocapsid is in the nonmembrane fraction of the cytoplasm of infected cells.

Cell Fractionation↗

PubMed1976

Respiratory syncytial virus-specific RNA synthesis in primary monkey kidney cell cultures.

Primary rhesus monkey kidney (MK) cell cultures were inoculated with respiratory syncytial virus and treated or untreated with actinomycin D before pulse labeling with uridine-5-3H. The virus-specific RNA synthesis was noted at its peak in the nucleoplasm and possibly less so in the cytoplasm of infected cells. At 48 and 72 hours post-inoculation (p.i.), small fractions of available cells were synthesising virus-specific RNA with labeling index of 15% and 18% respectively. By 48 hours p.i. syncytia started appearing and a higher grain count in the cytoplasm of actinomycin D-treated infected cells was noted.

Animals↗

PubMed1976

Maturation of ribosomes in yeast. I Kinetic analysis by labelling of high molecular weight rRNA species.

To study the maturation of ribosomes in Saccharomyces carlsbergensis, protoplasts were pulse labeled with [5-3H]uridine at 15 degrees C. Investigation of the cellular location of pulse-labelled ribosomal RNA precursor and mature ribosomal RNA shows that both the 37-S precursor RNA, common to both 17-S and 26-S rRNA, as well as the 29-S RNA, the direct precursor of 26-S rRNA, are located in the nucleus. Most of the 18-S RNA, the direct precursor of 17-S rRNA, is found in the cytoplasmic fraction. Apart from 37-S and 29-S RNA the nucleus also contains an appreciable amount of 26-S rRNA as well as a small quantity of 18-S RNA. These data indicate that processing of 29-S to 26-S RNA occurs in the nucleus, whereas the conversion of 18-S RNA to 17-S rRNA takes place in the cytoplasm. The kinetics of appearance of pulse-labelled 26-S and 17-S rRNA in the various cytoplasmic ribosomal particles indicate, that newly formed 40-S ribosomal particles are almost immediately incorporated into 80-S ribosomes and polysomes. On the other hand, there appears to exist a fairly large cytoplasmic pool of newly synthesized ribosomal particles containing 26-S rRNA and sedimenting at about 60 S. The kinetics of appearance of newly formed 26-S and 17-S rRNA in mature ribosomes show that the maturation of the large ribosomal subunit takes about twice as much time as that of the small subunit.

Cell Fractionation↗

PubMed1977

Synthesis of heterogeneous nuclear RNA in full-grown oocytes of Xenopus laevis (Daudin).

At various times following injection of either 3H-GTP or 32PO4 into full-grown (stage 6) Xenopus laevis oocytes, RNA has been extracted and fractionated on polyacrylamide gels. Based on size, base composition and incorporation data, we have defined the kinetics of synthesis and accumulation of ribosomal RNA (40S, 28S, 18S), heterogeneous RNA of high molecular weight (greater than 40S) and heterogeneous RNA migrating with molecular weights of from 4S to 40S. Nuclear isolations have been performed to determine the cellular distribution of these classes of RNA as a function of time. Evidence is presented which shows that stage 6 oocytes synthesize RNA which by virtue of its size, base composition, rapid turnover and nuclear location is equivalent to the heterogeneous nuclear RNA observed in somatic cells. In addition, the data suggest synthesis of a class of nuclear RNA with a half-life of several hours. A small fraction (5%) of the nuclear RNA is stable, enters the cytoplasm and may represent RNA added to the stockpile of maternal transcripts known to be present in stage 6 oocytes.

Animals↗

PubMed1975

RNA synthesis in cells infected with herpes simplex virus. IX. Evidence for accumulation of abundant symmetric transcripts in nuclei.

RNA extracted from nuclei of 8-h infected cells drove approximately 50% of herpes virus DNA into DNA-RNA hybrid. The same RNA, preannealed under conditions which allowed base pairing to take place, drove only 35% of the DNA into DNA-RNA hybrid; further annealing of the RNA did not diminish the amount of RNA sequences remaining available for subsequent hybridization with DNA. Upon denaturation of the preannealed RNA, the RNA sequences sequestered during preannealing became available again for hybridization with DNA. The base pairing that occurred during preincubation of the RNA was inter-molecular, since it was RNA concentration dependent and was not affected by limited alkaline hydrolysis. The nuclear viral transcripts that remained available for hybridization, after preannealing of the RNA, were subset of the RNA sequences that accumulated in the cytoplasm of infected cells. In addition, a small amount (derived from 5% or less of the viral DNA) of complementary transcripts was detected in the cytoplasm.

Base Sequence↗

PubMed1978

Cytology and quantitative cytochemistry of a poliferative atypical hemocytic condition in Mytilus edulis (Bivalvia, mollusca).

Proliferative lesions were found in 16 of 994 Mytilus edulis mussels obtained from the mouth of the River Lynher at Plymouth, England. These lesions were characterized by infiltration and replacement of the connective tissue by enlarged, atypical, mitotically active, basophilic, hemocyte-like cells. Cytologic examination indicated the involvement of two cell types in this disorder, both of which were similar in appearance to the much smaller, normal basophallic agranular hemocyte. The abnormal cells were rich in cytoplasmic RNA and had significantly higher DNA levels than normal hemocytes as determined by scanning microdensitometry. A few of the atypical cells contained small cytoplasmic granules that were positive for lysosomal hydrolases. This condition was observed sequentially from what is believed to be the earliest stage of the disease, in which only a few atypical cells were present, to the terminal stage, in which the connective tissue was almost completely replaced and the digestive gland cells were necrotized. Some potentially carcinogenic aromatic hydrocarbons have been identified in the substrate of the mussel bed by gas-liquid chromatography-mass spectrometry analysis.

Animals↗

PubMed1978

Extent of transcription of the E strand of polyoma virus DNA during the early phase of productive infection.

Early polyoma virus-specific RNA, in nuclei and cytoplasm of cells labeled with [(3)H]uridine, was analyzed by hybridization with filter-bound Hpa II fragments of polyoma DNA. About 40% of labeled cytoplasmic virus-specific RNA hybridized with Hpa II fragment 2, which represents about 40% of the region coding for E-strand mRNA's; less than 5% hybridized with fragments 1 or 3, which lie outside this region. A somewhat lower proportion (about 30%) of labeled nuclear virus-specific RNA hybridized with fragment 2, and a small but significant fraction (7 to 14%) hybridized with fragments 1 and 3. About two-thirds of the nuclear RNA which hybridized to fragment 1 was complementary to the E strand, and one-third was complementary to the L strand. Results did not vary greatly in samples labeled for periods of from 15 min to 3 h. The major species of pulse-labeled nuclear polyoma-specific RNA sedimented at 22S and thus is slightly larger than the 19S cytoplasmic mRNA. These results show that most early nuclear RNA ( approximately 75%) is transcribed from the region of the E strand, which codes for early mRNA's, and that there is probably a site at which transcription is terminated at the end of this region. However, a small amount of early nuclear RNA ( approximately 15%) is transcribed from the remainder of the E strand, perhaps by readthrough of this termination signal. In addition, there is a small amount of transcription from the L strand, whose significance is unclear. Neither the L-strand transcripts nor the nonmessenger E-strand transcripts are transported to the cytoplasm.

DNA, Viral↗

PubMed1979

Relationship between single-stranded DNA isolated from cultured muscular cells during differentiation and the transcription of messenger RNA.

Single-stranded DNA (ssDNA), equivalent to about 2% of the total nuclear DNA, was isolated by an improved method of hydroxyapatite chromatography from native nuclear DNA of rat myoblast cells and myotubes of the L6 line. Small quantities of 125I-labelled ssDNA were annealed with a large excess of unlabelled DNA, cytoplasmic RNA and mRNA from myoblasts or myotubes. The results indicated that ssDNA belongs to the non-repetitious portion of the cell genome and is formed of two distinct molecular fractions. The major ssDNA fractions (75%) consist of non-self-reassociating DNA sequences and the minor fraction (25%) consists of self-reassociating DNA sequences. About 30--32% and 25--26% of ssDNA from myoblast represent DNA sequences complementary to total cytplasmic RNAs and polyadenylated RNAs respectively. Hybridizations of ssDNA with an excess of RNA from myoblasts and/or myotubes show differences in the abundance and the diversity of mRNA during mascular differentiation. These differences were confirmed by DNA-driven reactions between 125I-labelled polyadenylated RNA and ssDNA in great excess.

Animals↗