Digital Genome-Wide ncRNA Expression, Including SnoRNAs, across 11 Human Tissues Using PolyA-Neutral Amplification

Non-coding RNAs (ncRNAs) are an essential class of molecular species that have been difficult to monitor on high throughput platforms due to frequent lack of polyadenylation. Using a polyadenylation-neutral amplification protocol and next-generation sequencing, we explore ncRNA expression in eleven human tissues. ncRNAs 7SL, U2, 7SK, and HBII-52 are expressed at levels far exceeding mRNAs. C/D and H/ACA box snoRNAs are associated with rRNA methylation and pseudouridylation, respectively: spleen expresses both, hypothalamus expresses mainly C/D box snoRNAs, and testes show enriched expression of both H/ACA box snoRNAs and RNA telomerase TERC. Within the snoRNA 14q cluster, 14q(I-6) is expressed at much higher levels than other cluster members. More reads align to mitochondrial than nuclear tRNAs. Many lincRNAs are actively transcribed, particularly those overlapping known ncRNAs. Within the Prader-Willi syndrome loci, the snoRNA HBII-85 (group I) cluster is highly expressed in hypothalamus, greater than in other tissues and greater than group II or III. Additionally, within the disease locus we find novel transcription across a 400,000 nt span in ovaries. This genome-wide polyA-neutral expression compendium demonstrates the richness of ncRNA expression, their high expression patterns, their function-specific expression patterns, and is publicly available.

[1]  J. Rinn,et al.  Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression , 2009, Proceedings of the National Academy of Sciences.

[2]  K. Collins,et al.  Telomerase RNA level limits telomere maintenance in X-linked dyskeratosis congenita. , 2006, Genes & development.

[3]  W. Keith,et al.  Telomerase redefined: integrated regulation of hTR and hTERT for telomere maintenance and telomerase activity. , 2008, Biochimie.

[4]  Jeannie T. Lee,et al.  Tsix, a gene antisense to Xist at the X-inactivation centre , 1999, Nature Genetics.

[5]  Michael Thomas,et al.  MALAT-1, a novel noncoding RNA, and thymosin β4 predict metastasis and survival in early-stage non-small cell lung cancer , 2003, Oncogene.

[6]  Laurent Lestrade,et al.  snoRNA-LBME-db, a comprehensive database of human H/ACA and C/D box snoRNAs , 2005, Nucleic Acids Res..

[7]  Sean R. Eddy,et al.  Rfam: an RNA family database , 2003, Nucleic Acids Res..

[8]  U. Meier,et al.  The many facets of H/ACA ribonucleoproteins , 2005, Chromosoma.

[9]  J. Sutcliffe,et al.  Primate brain-specific cytoplasmic transcript of the Alu repeat family , 1987, Molecular and cellular biology.

[10]  G. Chrousos,et al.  Noncoding RNA Gas5 Is a Growth Arrest– and Starvation-Associated Repressor of the Glucocorticoid Receptor , 2010, Science Signaling.

[11]  R. Nicholls,et al.  Genome organization, function, and imprinting in Prader-Willi and Angelman syndromes. , 2001, Annual review of genomics and human genetics.

[12]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[13]  E. Rocha Codon usage bias from tRNA's point of view: redundancy, specialization, and efficient decoding for translation optimization. , 2004, Genome research.

[14]  A. Hüttenhofer,et al.  Identification of brain-specific and imprinted small nucleolar RNA genes exhibiting an unusual genomic organization. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[15]  David Haussler,et al.  The UCSC Genome Browser database: update 2010 , 2009, Nucleic Acids Res..

[16]  A. Ferré-D’Amaré,et al.  The Box H/ACA Ribonucleoprotein Complex: Interplay of RNA and Protein Structures in Post-transcriptional RNA Modification* , 2009, The Journal of Biological Chemistry.

[17]  Carolyn J. Brown,et al.  The human XIST gene: Analysis of a 17 kb inactive X-specific RNA that contains conserved repeats and is highly localized within the nucleus , 1992, Cell.

[18]  Mary Goldman,et al.  The UCSC Genome Browser database: update 2011 , 2010, Nucleic Acids Res..

[19]  M. Gross,et al.  The human ASM (adult skeletal muscle) gene: expression and chromosomal assignment to 11p15. , 1991, Biochemical and biophysical research communications.

[20]  Ronghua Chen,et al.  Digital transcriptome profiling using selective hexamer priming for cDNA synthesis , 2009, Nature Methods.

[21]  Tao Pan,et al.  Tissue-Specific Differences in Human Transfer RNA Expression , 2006, PLoS genetics.

[22]  M. W. Gray,et al.  Pseudouridine in RNA: What, Where, How, and Why , 2000, IUBMB life.

[23]  M. Blasco,et al.  Telomerase regulation and stem cell behaviour. , 2006, Current opinion in cell biology.

[24]  Marwan Shinawi,et al.  Prader-Willi phenotype caused by paternal deficiency for the HBII-85 C/D box small nucleolar RNA cluster , 2008, Nature Genetics.

[25]  Martina Paulsen,et al.  Identification of tandemly-repeated C/D snoRNA genes at the imprinted human 14q32 domain reminiscent of those at the Prader-Willi/Angelman syndrome region. , 2002, Human molecular genetics.

[26]  A. Hüttenhofer,et al.  The expanding snoRNA world. , 2002, Biochimie.

[27]  Ronghua Chen,et al.  DNA copy number, including telomeres and mitochondria, assayed using next-generation sequencing , 2010, BMC Genomics.

[28]  G. Daley,et al.  Telomere elongation in induced pluripotent stem cells from dyskeratosis congenita patients , 2009, Nature.

[29]  John N. Hutchinson,et al.  A screen for nuclear transcripts identifies two linked noncoding RNAs associated with SC35 splicing domains , 2007, BMC Genomics.

[30]  S. Eddy Non–coding RNA genes and the modern RNA world , 2001, Nature Reviews Genetics.

[31]  John N. Hutchinson,et al.  An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles. , 2009, Molecular cell.

[32]  T. Krude,et al.  Noncoding human Y RNAs are overexpressed in tumours and required for cell proliferation , 2008, British Journal of Cancer.

[33]  J. Mattick,et al.  Non-coding RNA. , 2006, Human molecular genetics.

[34]  Ting Wang,et al.  The UCSC Genome Browser Database: update 2009 , 2008, Nucleic Acids Res..

[35]  Christopher J. Lee,et al.  A transcriptional sketch of a primary human breast cancer by 454 deep sequencing , 2009, BMC Genomics.

[36]  David I. Smith,et al.  Long, abundantly expressed non-coding transcripts are altered in cancer. , 2008, Human molecular genetics.