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At least 253 records · Page 14Linked to original sources

In vitro transcription of two Epstein-Barr virus specified small RNA molecules.

Cloned DNA from the EcoRI J fragment of EBV has been used as template for in vitro transcription experiments using cell-free extracts prepared from HeLa or KB cells. Two EBV specific RNAs each about 175 bases in length were synthesised and nuclease 51 mapping experiments determined that these in vitro products corresponded precisely to the in vivo species obtained from Raji cells. These two RNA molecules are transcribed by RNA polymerase III and in common with other pol III-synthesised RNAs the coding sequences contain intragenic control regions. The relative abundance of the two RNAs synthesised in vitro differs from that observed in vivo.

Base Sequence↗

Copy-number of broad host-range plasmid R1162 is regulated by a small RNA.

We have shown previously [Kim, K. and Meyer, R.J. (1985) J. Mol. Biol. 185,755-767] that copy-number of the broad host-range plasmid R1162 is controlled by the amounts of two proteins, encoded by cotranscribed genes comprising a region of the plasmid called RepI. We have now demonstrated that expression of RepI is negatively regulated by a 75 base RNA that is complementary to a segment of the RepI message. Increased intracellular amounts of RNA molecules that include this segment relieve the inhibition of RepI gene expression, suggesting that the target for regulation is the mRNA itself. A mutation decreasing the amount of the 75 base RNA results in elevated plasmid copy-number. Thus, consistent with our previous observations, regulation of the expression of the RepI genes is a factor in controlling plasmid copy-number.

Base Sequence↗

Single base bulges in small RNA hairpins enhance ethidium binding and promote an allosteric transition.

A set of four RNA hairpin helices has been prepared by in vitro transcription with T7 RNA polymerase. The hairpins all contain the same nine base pair helix, but with an extra A, C, or U residue forming a bulge at one position; the fourth hairpin is a perfect helix with no bulge. The helix with a bulged A duplicates six base pairs of a helix in the 16S rRNA known to have an unusually high affinity for ethidium bromide [J. M. Kean, S. A. White, and D. E. Draper, Biochemistry 24, 5062 (1985)]. Binding and chemical cleavage studies with ethidium or the reagent methidiumpropylEDTA-Fe(II) [MPE-Fe(II)] showed that the sequence CpG is a preferred intercalation site whether or not a bulge is present; all three bulged bases enhance intercalation at the CpG sequence by an order of magnitude; and intercalation in a bulged helix results in a concerted conformational change involving the entire helix backbone, again dependent on the presence of a bulge but independent of the particular base. These results suggest that an extra sugar-phosphate residue in an RNA helix backbone has a dramatic effect on the ability of the RNA to adopt new conformations. This effect could be an important reason for the conservation of bulges at certain positions in ribosomal and other RNAs.

Allosteric Site↗

Synthesis and behaviour of small RNA species of CHO cells submitted to a heat chick.

Incubation of Chinese hamster ovary (CHO) cells for 1 hour at 43 degrees C results in an inhibition of high molecular weight RNA synthesis while most of the ow molecular weight RNAs are still synthesized. In cells returned to 37 degrees C, the transcription of high molecular weight RNA is reinitiated after 7 h recovery. The synthesis of snRNA A, C, D which are transcribed by RNA polymerase B (II) is inhibited in cells incubated at 43 degrees C while the synthesis of 4S, 5S, L and K components is not affected. During the recovery period the synthesis of low molecular weight RNA is increased variously according to the components relative to control cells : x 1.5 for 5S RNA to x 8 for snK. After 9 h recovery at 37 degrees C, snA and SnD are again synthesized but newly synthesized snC does not appear in the nucleus while a putative preC component accumulates in the cytoplasm. On the other hand, the distribution of all the pre-existing low molecular weight RNAs is not affected by the heat shock.

Animals↗

Serological relations between twelve small RNA viruses of insects.

Serological tests were done to examine the relationships between twelve picorna-like viruses of insects. The results of the tests indicated that the majority of the viruses are unrelated. However, cricket paralysis virus, isolated from Australian wild field crickets, appeared to be identical to Drosophila C virus, independently isolated in France. Cricket paralysis virus was infective for adults of Drosophila melanogaster and its infectivity towards Galleria melonella was neutralised by Drosophila C virus antiserum. It is therefore concluded that cricket paralysis virus and Drosophila C virus are very closely related if not identical.

Animals↗

Small RNA helices as substrates for aminoacylation and their relationship to charging of transfer RNAs.

RNA microhelices that reconstruct the acceptor stems of transfer RNAs can be aminoacylated. The anticodon-independent aminoacylation is sequence-specific and suggests a relationship between amino acids and nucleotide sequences which is different from that of the classical genetic code. The specific aminoacylation of RNA microhelices also suggests a highly differentiated adaptation of the structures of aminoacyl-tRNA synthetases to sequences in the acceptor stems of transfer RNAs.

Amino Acid Sequence↗

Effect of RyhB small RNA on global iron use in Escherichia coli.

RyhB is a noncoding RNA regulated by the Fur repressor. It has previously been shown to cause the rapid degradation of a number of mRNAs that encode proteins that utilize iron. Here we examine the effect of ectopic RyhB production on global gene expression by microarray analysis. Many of the previously identified targets were found, as well as other mRNAs encoding iron-binding proteins, bringing the total number of regulated operons to at least 18, encoding 56 genes. The two major operons involved in Fe-S cluster assembly showed different behavior; the isc operon appears to be a direct target of RyhB action, while the suf operon does not. This is consistent with previous findings suggesting that the suf genes but not the isc genes are important for Fe-S cluster synthesis under iron-limiting conditions, presumably for essential iron-binding proteins. In addition, we observed repression of Fur-regulated genes upon RyhB expression, interpreted as due to intracellular iron sparing resulting from reduced synthesis of iron-binding proteins. Our results demonstrate the broad effects of a single noncoding RNA on iron homeostasis.

Bacterial Proteins↗

Origin of small RNA in von Magnus particles of influenza virus.

A clone of recombinant virus obtained from the cross between WSN and Hong Kong strains of influenza virus gave rise to progeny containing predominantly von Magnus particles. In the electropherogram of virus RNA, the P3 gene was markedly diminished, and a new species of RNA (extra RNA) was present in addition to eight gene segments. The origin of the extra RNA was studied by two-dimensional gel electrophoresis of T1 RNase-generated oligonucleotides. Four out of five large oligonucleotide spots present in the extra RNA matched to those contained by the P3 gene. It was concluded that the extra RNA was derived from the P3 gene probably by deletion. The possible origin of the spot which was present in the extra RNA but not in eight gene segments including P3 was discussed.

Defective Viruses↗

Isolation of the ends of La Crosse virus small RNA as a double-stranded structure.

The genome of La Crosse virus, a member of the Bunyaviridae, is made up of three molecules. Circular nucleocapsid structures, in three size classes, have been isolated from La Crosse virus (Obijeski et al. J. Virol. 20:664-675, 1976). Recently, Obijeski et al. (Nucleic Acids Res. 8:2431-2438) have found that the 5' and 3' ends of each segment are complementary in sequence. We determined that a 5' and 3' end complementary structure, predicted by the rules of Tinoco et al. (Nature [London] 230:362-367), can and will anneal under certain conditions. This structure is resistant to RNase in high-salt medium and can be isolated in a reasonably high yield.

Base Sequence↗

Epstein-Barr virus small RNA (EBER) genes: differential regulation during lytic viral replication.

In every latently Epstein-Barr virus-infected cell the viral genes EBER-1 and EBER-2 are transcribed by polymerase III. In lytically infected cells in vivo the EBER genes could not be detected. However, in cell culture downregulation could not be confirmed, and hence the relevance of this shutdown to the replication of the virus was not clear. We assayed the transcriptional activity of the EBER genes by nuclear run-on assays with enriched lytically infected cells and demonstrated that EBER-1 and EBER-2 are differentially downregulated on the transcriptional level during the switch to lytic viral replication. This downregulation was an early event during the lytic replication of the virus.

Base Sequence↗

La antigen recognizes and binds to the 3'-oligouridylate tail of a small RNA.

The La antigen is a cellular protein which interacts with many RNA species that are products of RNA polymerase III, including the adenovirus virus-associated (VA) RNAs. We demonstrate that the efficiency of antigen binding in vitro is determined by the number of U residues at the RNA 3' terminus. Forms of VA RNAI with more than two terminal U residues are fully bound, forms with two U residues are partially bound, and forms with fewer than two U residues are not bound at all. The antigen can be covalently linked to VA RNA by UV irradiation, and the site of cross-linking is shown to contain the 3' terminus of the RNA. We conclude that the antigen recognizes the U-rich 3' tail of VA RNA, and presumably that of other polymerase III products, and that it binds at or close to this site.

Adenine Nucleotides↗

Physicochemical characterization of a small RNA virus associated with baculovirus infection in Trichoplusia ni.

An RNA virus was isolated from Trichoplusia ni larvae infected with inoculum of the baculovirus of Autographa californica. Physicochemical characterization has revealed a 35-nm virus with a density in CsCl of 1.3 g/cm3 and a sedimentation coefficient of 200S. The virus consisted of 10--15% single-stranded RNA with an apparent molecular weight of 1.9 x 10(6) and coat protein with a subunit size of 67,000. The virus is similar to the Nudaurelia capensis beta virus and also possesses some interesting similarities to the mammalian caliciviruses. The proposed cryptogram of the virus is R/1:1.9/12:S/S:I/O.

Animals↗

Naturally occurring IgM antibodies to a small RNA insect virus in some mammalian sera in New Zealand.

Antibodies to cricket paralysis virus were demonstrated in sera from a pig, a horse, and numerous cattle in New Zealand. The reactions in immunodiffusion tests were variable, but significant and consistent reactions were obtained with these sere in virus neutralization assays using Drosophila cells. The sedimentation coefficient of the antibodies was 19S, and their activity was destroyed by treatment with 0.1 M 2-mercaptoethanol, indicating that they were of the IgM class.

Animals↗