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2013 ; 14
(5
): R45
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Insights into snoRNA biogenesis and processing from PAR-CLIP of snoRNA core
proteins and small RNA sequencing
#MMPMID23706177
Kishore S
; Gruber AR
; Jedlinski DJ
; Syed AP
; Jorjani H
; Zavolan M
Genome Biol
2013[May]; 14
(5
): R45
PMID23706177
show ga
BACKGROUND: In recent years, a variety of small RNAs derived from other RNAs with
well-known functions such as tRNAs and snoRNAs, have been identified. The
functional relevance of these RNAs is largely unknown. To gain insight into the
complexity of snoRNA processing and the functional relevance of snoRNA-derived
small RNAs, we sequence long and short RNAs, small RNAs that co-precipitate with
the Argonaute 2 protein and RNA fragments obtained in photoreactive
nucleotide-enhanced crosslinking and immunoprecipitation (PAR-CLIP) of core
snoRNA-associated proteins. RESULTS: Analysis of these data sets reveals that
many loci in the human genome reproducibly give rise to C/D box-like snoRNAs,
whose expression and evolutionary conservation are typically less pronounced
relative to the snoRNAs that are currently cataloged. We further find that
virtually all C/D box snoRNAs are specifically processed inside the regions of
terminal complementarity, retaining in the mature form only 4-5 nucleotides
upstream of the C box and 2-5 nucleotides downstream of the D box. Sequencing of
the total and Argonaute 2-associated populations of small RNAs reveals that
despite their cellular abundance, C/D box-derived small RNAs are not efficiently
incorporated into the Ago2 protein. CONCLUSIONS: We conclude that the human
genome encodes a large number of snoRNAs that are processed along the canonical
pathway and expressed at relatively low levels. Generation of snoRNA-derived
processing products with alternative, particularly miRNA-like, functions appears
to be uncommon.
|Argonaute Proteins/*metabolism
[MESH]
|Cross-Linking Reagents/metabolism
[MESH]
|Genome, Human
[MESH]
|HEK293 Cells
[MESH]
|HeLa Cells
[MESH]
|Humans
[MESH]
|Immunoprecipitation
[MESH]
|Models, Molecular
[MESH]
|Molecular Sequence Data
[MESH]
|RNA, Small Nucleolar/*analysis/metabolism
[MESH]
|Ribonucleoproteins, Small Nucleolar/*analysis
[MESH]