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Effect of the deletion of the reported ERH binding site embedded in the DGCR8 N-terminus and/or global ERH knockdown on the expression of mature miRNAs

ABSTRACT: MicroRNAs are small non-coding RNAs that mediate post-transcriptional silencing of most mammalian genes. They are generated in a multi-step process initiated by the Microprocessor, a protein complex composed of DROSHA and DGCR8. Recent studies have described the phenomenon of “cluster assistance”, in which a prototypic primary miRNA hairpin can license the Microprocessor-mediated processing of a clustered suboptimal hairpin in cis. Mechanistic analyses led to the identification of ERH (enhancer of rudimentary homolog) as a critical factor in this process. Notably, ERH has been demonstrated to associate with the N-terminus of DGCR8, but it has not been demonstrated that this complex is required for cluster assistance. Here, we have used prime editing to generate alleles that encode a DGCR8 Δ103-126Q mutant that cannot bind ERH anymore. In contrast to global ERH knockdown, disruption of the ERH binding motif in DGCR8 affects processing of only a subset of cluster assistance-unrelated pri-miRNAs. Thus, ERH-mediated cluster assistance is independent of its described association with DGCR8, indicating that ERH confers two distinct roles in primary miRNA biogenesis: One driven by its direct binding to the Microprocessor via DGCR8 and the other as a DGCR8/ERH complex-independent factor in cluster assistance.

ORGANISM(S): Homo sapiens

PROVIDER: GSE311273 | GEO | 2025/12/05

REPOSITORIES: GEO

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