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Small nucleolar RNAs encoded by introns of the human cell cycle regulatory gene RCC1.

ABSTRACT: Eukaryotic cells contain a large number of U small nuclear RNAs (U-snRNAs) involved in various RNA processing reactions in the nucleoplasm and nucleous. Most of the U-snRNAs have 5'-terminal caps added to the end of the primary transcript. Here we describe two variants of a snRNA, called U17, identified in human HeLa cells. U17 RNA may be involved in ribosome biogenesis since it is found in the nucleolus and sediments with 40S structures possibly representing nascent ribosomal subunits. U17 RNAs contain no cap but have a monophosphate at the 5'-terminus indicating that they are processed from longer precursors. The U17 RNAs are encoded within introns 1 and 2 of the single copy gene RCC1 which codes for an important cell cycle regulatory protein. In HeLa cell S-100 extract, U17 RNA is faithfully excised from a longer RNA transcript derived from the intron yielding 5'-monophosphorylated RNA. These data suggest that U17 RNAs are not independently transcribed but are processed out of the RCC1 pre-mRNA or out of the spliced introns.

SUBMITTER: Kiss T

PROVIDER: S-EPMC413544 | biostudies-other | 1993 Jul

REPOSITORIES: biostudies-other

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Small nucleolar RNAs encoded by introns of the human cell cycle regulatory gene RCC1.

Kiss T T Filipowicz W W

The EMBO journal 19930701 7

Eukaryotic cells contain a large number of U small nuclear RNAs (U-snRNAs) involved in various RNA processing reactions in the nucleoplasm and nucleous. Most of the U-snRNAs have 5'-terminal caps added to the end of the primary transcript. Here we describe two variants of a snRNA, called U17, identified in human HeLa cells. U17 RNA may be involved in ribosome biogenesis since it is found in the nucleolus and sediments with 40S structures possibly representing nascent ribosomal subunits. U17 RNAs ...[more]

PMID: 8335005

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Project description:Purpose: Small regulatory RNAs (sRNAs) are short transcripts that base-pair to mRNA targets or interact with regulatory proteins. sRNA function has been extensively studied in Gram-negative bacteria; comparatively less is known about sRNAs in Firmicutes. Here we investigate two sRNAs encoded by the virulence plasmid pXO1 of Bacillus anthracis, the causative agent of anthrax. We designated the sRNAs as as “XrrA” and “XrrB” (for pXO1-encoded regulatory RNA). We constructed deletion mutants ∆xrrA, ∆xrrB, and ∆xrrA∆xrrB in an Ames strain background and compared gene expression of these mutants to that of the Ames-derived parent strain using RNA-seq. Methods: Total RNA from Ames strains UTA37 (Parent), UTA38 (∆xrrB), UTA39 (∆xrrA), UTA41 (∆xrrA∆xrrB) was isolated from cultures grown in CA-CO2 in triplicate, using saturated phenol: chloroform extraction. One microgram of pure and quality-checked RNA from each sample was subjected to Illumina sequencing by synthesis using a NextSeq550 sequencer. Sequencing by synthesis was performed to generate 75bp paired-end reads. Two 130M-read sequencing runs (Run1 and Run2) were performed and an average of 31M reads per sample were obtained. Results: All bioinformatic analysis was performed using the publicly available Galaxy web resource (https://usegalaxy.org/). Comparison of the transcriptomes of a virulent Ames-derived strain to isogenic sRNA-null mutants revealed multiple 4.0- to >100-fold differences in gene expression. Deletion of xrrA resulted in 50 transcripts showing a ≥ 4.0-fold change in expression compared to the parent strain. In contrast, deletion of xrrB led to one transcript having a ≥ 4.0-fold change in expression compared to the parent strain. In the ∆xrrA∆xrrB mutant, levels of 116 transcripts were affected with a fold-change of ≥ 4.0. Many sRNA-regulated targets were chromosome genes associated with branched-chain amino acid metabolism, proteolysis, and transmembrane transport. Conclusions: In this work, we provide experimental evidence for sRNA-mediated regulation in the mammalian pathogen B. anthracis. XrrA and XrrB are the first regulatory RNAs examined in B. anthracis and are shown here to have multiple effects on gene expression in this pathogen. The data suggest distinct regulatory roles for XrrA, as well as some functional overlap between XrrA and XrrB.

Dataset Information

Small nucleolar RNAs encoded by introns of the human cell cycle regulatory gene RCC1.

Publications

Small nucleolar RNAs encoded by introns of the human cell cycle regulatory gene RCC1.

Similar Datasets

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