Dataset Information

Gene expression profiling in THP1 cells after PAPD5 or GLD2 knockdown

ABSTRACT: Next-generation sequencing experiments have shown that microRNAs are expressed in many different isoforms (isomiRs), whose biological relevance is often unclear. We found that mature miR-21, the most widely researched microRNA because of its importance in human disease, is produced in two prevalent isomiR forms that differ by one nucleotide at their 3’ end, and moreover that the 3’ end of miR-21 is post transcriptionally adenylated by the noncanonical poly(A) polymerase PAPD5. PAPD5 knockdown caused an increase in the miR-21 expression level, suggesting that PAPD5-mediated adenylation of miR-21 leads to its degradation. Exoribonuclease knockdown experiments followed by small RNA sequencing suggested that PARN degrades miR-21 in the 3’-to-5’ direction. In accordance with this model, microarray expression profiling demonstrated that PAPD5 knockdown results in a downregulation of miR-21 target mRNAs. We found that disruption of the miR-21 adenylation and degradation pathway is a general feature in tumors across a wide range of tissues, as evidenced by data from The Cancer Genome Atlas, as well as in the non-cancerous proliferative disease psoriasis. We conclude that PAPD5 and PARN mediate degradation of oncomiR miR-21 through a tailing and trimming process, and that this pathway is disrupted in cancer and other proliferative diseases.

ORGANISM(S): Homo sapiens

PROVIDER: GSE57939 | GEO | 2014/05/24

SECONDARY ACCESSION(S): PRJNA248456

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Mutations in genes encoding components of the telomerase holoenzyme complex result in a spectrum of rare genetic disorders known as telomere diseases, including dyskeratosis congenita (DC). A consistent finding in DC due to pathogenic mutations in DKC1, which encodes dyskerin, is decreased steady-state levels of the non-coding RNA component of telomerase (TERC) and thus impaired telomere maintenance. Dyskerin binds hundreds of other small nucleolar RNAs (snoRNAs). However, the mechanisms by which DKC1 mutations cause variable impacts on these snoRNAs are poorly understood, which is a barrier to understanding disease mechanisms in DC beyond impaired telomere maintenance. Here, using somatic and induced pluripotent stem cells (iPSCs) from DC patients with DKC1 mutations and CRISPR-Cas9-engineered iPSCs, we show that mutations in the N-terminal extension domain (NTE) of dyskerin dysregulate the biogenesis of a subset of snoRNAs, with the most prominent effect on scaRNA13 (small Cajal body-specific RNA 13). In patient iPSCs carrying the del37L dyskerin NTE-domain mutation but not in those with C-terminal mutations, nascent scaRNA13 transcripts showed a discrete population of 3´-extended forms, as seen in the setting of DC-causing mutations in the PARN (polyA-specific ribonuclease) gene. By deep sequencing of RNA 3´ ends, we found that aberrant scaRNA13 transcripts were composed of genomically-encoded extensions and post-transcriptionally oligoadenylated species, mediated by the noncanonical polymerase PAPD5, which counters PARN. NTE domain mutations generated using CRISPR-Cas9 engineering of the endogenous DKC1 recapitulated the scaRNA13 3´-end processing defects seen in del37L patient cells. Conversely, repair of the DKC1 del37L mutation and genetic or pharmacological manipulation of PAPD5 rescued scaRNA13 end processing defects and steady-state levels. Analysis of the human telomerase cryo-EM structure showed that the dyskerin NTE interacts with 3´ end of bound RNA, suggesting that mutations in this domain impair 3´ end protection of nascent scaRNA13 in addition to canonical functions in snoRNA stabilization. Our results provide mechanistic insights into the interplay of dyskerin and the PARN/PAPD5 axis in the biogenesis and accumulation of snoRNAs beyond TERC, which has important implications for our broader understanding of ncRNA dysregulation in human diseases.

Dataset Information

Gene expression profiling in THP1 cells after PAPD5 or GLD2 knockdown

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure