Sequencing of Argonaute-bound microRNA/mRNA hybrids reveals regulation of the unfolded protein response by microRNA-320a. PLoS Genet 2021 Dec;17(12):e1009934
Date
12/17/2021Pubmed ID
34914716Pubmed Central ID
PMC8675727DOI
10.1371/journal.pgen.1009934Scopus ID
2-s2.0-85121763186 (requires institutional sign-in at Scopus site) 15 CitationsAbstract
MicroRNAs (miRNA) are short non-coding RNAs widely implicated in gene regulation. Most metazoan miRNAs utilize the RNase III enzymes Drosha and Dicer for biogenesis. One notable exception is the RNA polymerase II transcription start sites (TSS) miRNAs whose biogenesis does not require Drosha. The functional importance of the TSS-miRNA biogenesis is uncertain. To better understand the function of TSS-miRNAs, we applied a modified Crosslinking, Ligation, and Sequencing of Hybrids on Argonaute (AGO-qCLASH) to identify the targets for TSS-miRNAs in HCT116 colorectal cancer cells with or without DROSHA knockout. We observed that miR-320a hybrids dominate in TSS-miRNA hybrids identified by AGO-qCLASH. Targets for miR-320a are enriched for the eIF2 signaling pathway, a downstream component of the unfolded protein response. Consistently, in miR-320a mimic- and antagomir- transfected cells, differentially expressed gene products are associated with eIF2 signaling. Within the AGO-qCLASH data, we identified the endoplasmic reticulum (ER) chaperone calnexin as a direct miR-320a down-regulated target, thus connecting miR-320a to the unfolded protein response. During ER stress, but not amino acid deprivation, miR-320a up-regulates ATF4, a critical transcription factor for resolving ER stress. In summary, our study investigates the targetome of the TSS-miRNAs in colorectal cancer cells and establishes miR-320a as a regulator of unfolded protein response.
Author List
Fields CJ, Li L, Hiers NM, Li T, Sheng P, Huda T, Shan J, Gay L, Gu T, Bian J, Kilberg MS, Renne R, Xie MAuthor
Tongjun Gu PhD Associate Investigator in the Hematopoiesis & Stem Cell Biology department at BloodCenter of WisconsinMESH terms used to index this publication - Major topics in bold
Activating Transcription Factor 4Antagomirs
Argonaute Proteins
Calnexin
Cell Movement
Cell Proliferation
Colorectal Neoplasms
DEAD-box RNA Helicases
Endoplasmic Reticulum
Endoplasmic Reticulum Stress
Eukaryotic Initiation Factor-2
Gene Knockout Techniques
HCT116 Cells
Humans
MicroRNAs
Ribonuclease III
Signal Transduction
Transcription Initiation Site
