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Structure paper

TitleThe structural landscape of Microprocessor-mediated processing of pri-let-7 miRNAs.
Journal, issue, pagesMol Cell, Vol. 84, Issue 21, Page 4175-44190.e6, Year 2024
Publish dateNov 7, 2024
AuthorsAnkur Garg / Renfu Shang / Todor Cvetanovic / Eric C Lai / Leemor Joshua-Tor /
PubMed AbstractMicroRNA (miRNA) biogenesis is initiated upon cleavage of a primary miRNA (pri-miRNA) hairpin by the Microprocessor (MP), composed of the Drosha RNase III enzyme and its partner DGCR8. Multiple pri- ...MicroRNA (miRNA) biogenesis is initiated upon cleavage of a primary miRNA (pri-miRNA) hairpin by the Microprocessor (MP), composed of the Drosha RNase III enzyme and its partner DGCR8. Multiple pri-miRNA sequence motifs affect MP recognition, fidelity, and efficiency. Here, we performed cryoelectron microscopy (cryo-EM) and biochemical studies of several let-7 family pri-miRNAs in complex with human MP. We show that MP has the structural plasticity to accommodate a range of pri-miRNAs. These structures revealed key features of the 5' UG sequence motif, more comprehensively represented as the "flipped U with paired N" (fUN) motif. Our analysis explains how cleavage of class-II pri-let-7 members harboring a bulged nucleotide generates a non-canonical precursor with a 1-nt 3' overhang. Finally, the MP-SRSF3-pri-let-7f1 structure reveals how SRSF3 contributes to MP fidelity by interacting with the CNNC motif and Drosha's Piwi/Argonaute/Zwille (PAZ)-like domain. Overall, this study sheds light on the mechanisms for flexible recognition, accurate cleavage, and regulated processing of different pri-miRNAs by MP.
External linksMol Cell / PubMed:39368465 / PubMed Central
MethodsEM (single particle)
Resolution2.8 - 3.2 Å
Structure data

43819

9asm

EMDB-43819, PDB-9asm:
Human Drosha and DGCR8 in complex with Pri-let-7f1
Method: EM (single particle) / Resolution: 2.8 Å

43820

9asn

EMDB-43820, PDB-9asn:
Human Drosha and DGCR8 in complex with Pri-miR-98
Method: EM (single particle) / Resolution: 3.2 Å

43821

9aso

EMDB-43821, PDB-9aso:
Human Drosha and DGCR8 in complex with Pri-let-7a2
Method: EM (single particle) / Resolution: 2.9 Å

43822

9asp

EMDB-43822, PDB-9asp:
Human Drosha and DGCR8 in complex with Pri-let-7a1
Method: EM (single particle) / Resolution: 3.2 Å

43823

9asq

EMDB-43823, PDB-9asq:
Human Drosha, DGCR8 and SRSF3 in complex with Pri-let-7f1
Method: EM (single particle) / Resolution: 3.0 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-CA:
Unknown entry

ChemComp-HOH:
WATER

Source
  • homo sapiens (human)
KeywordsRNA BINDING PROTEIN/RNA / RNAi / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA complex

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