RRC ID 76084
Author Vieux KF, Prothro KP, Kelley LH, Palmer C, Maine EM, Veksler-Lublinsky I, McJunkin K.
Title Screening by deep sequencing reveals mediators of microRNA tailing in C. elegans.
Journal Nucleic Acids Res
Abstract microRNAs are frequently modified by addition of untemplated nucleotides to the 3' end, but the role of this tailing is often unclear. Here we characterize the prevalence and functional consequences of microRNA tailing in vivo, using Caenorhabditis elegans. MicroRNA tailing in C. elegans consists mostly of mono-uridylation of mature microRNA species, with rarer mono-adenylation which is likely added to microRNA precursors. Through a targeted RNAi screen, we discover that the TUT4/TUT7 gene family member CID-1/CDE-1/PUP-1 is required for uridylation, whereas the GLD2 gene family member F31C3.2-here named GLD-2-related 2 (GLDR-2)-is required for adenylation. Thus, the TUT4/TUT7 and GLD2 gene families have broadly conserved roles in miRNA modification. We specifically examine the role of tailing in microRNA turnover. We determine half-lives of microRNAs after acute inactivation of microRNA biogenesis, revealing that half-lives are generally long (median = 20.7 h), as observed in other systems. Although we observe that the proportion of tailed species increases over time after biogenesis, disrupting tailing does not alter microRNA decay. Thus, tailing is not a global regulator of decay in C. elegans. Nonetheless, by identifying the responsible enzymes, this study lays the groundwork to explore whether tailing plays more specialized context- or miRNA-specific regulatory roles.
Volume 49(19)
Pages 11167-11180
Published 2021-11-8
DOI 10.1093/nar/gkab840
PII 6377831
PMID 34586415
PMC PMC8565341
MeSH Adenosine Monophosphate / metabolism Animals Caenorhabditis elegans / classification Caenorhabditis elegans / genetics* Caenorhabditis elegans / metabolism Caenorhabditis elegans Proteins / genetics* Caenorhabditis elegans Proteins / metabolism Cell Cycle Proteins / genetics* Cell Cycle Proteins / metabolism Chickens / classification Chickens / genetics Chickens / metabolism Conserved Sequence Gene Expression Regulation Genome, Helminth* Half-Life Humans Mice MicroRNAs / antagonists & inhibitors MicroRNAs / classification MicroRNAs / genetics* MicroRNAs / metabolism Phylogeny RNA Interference RNA Stability RNA, Helminth / classification RNA, Helminth / genetics* RNA, Helminth / metabolism Species Specificity Uridine Monophosphate / metabolism* Zebrafish / classification Zebrafish / genetics Zebrafish / metabolism
Resource
C.elegans tm1021 tm4344