Regulation of Microprocessor assembly and localization via Pasha’s WW domain inC. elegans

Author:

Montgomery Brooke E.,Knittel Thiago L.ORCID,Reed Kailee J.,Chong Madeleine C.,Isolehto Ida J.,Cafferty Erin R.,Smith Margaret J.,Sprister Reese A.,Magelky Colin N.,Scherman Hataichanok,Ketting Rene F.,Montgomery Taiowa A.ORCID

Abstract

ABSTRACTPrimary microRNA (pri-miRNA) transcripts are processed by the Microprocessor, a protein complex that includes the ribonuclease Drosha and its RNA binding partner DGCR8/Pasha. We developed a live, whole animal, fluorescence-based sensor that reliably monitors pri-miRNA processing with high sensitivity inC. elegans. Through a forward genetic selection for alleles that desilence the sensor, we identified a mutation in the conserved G residue adjacent to the namesake W residue of Pasha’s WW domain. Using genome editing we also mutated the W residue and reveal that both the G and W residue are required for dimerization of Pasha and proper assembly of the Microprocessor. Surprisingly, we find that the WW domain also facilitates nuclear localization of Pasha, which in turn promotes nuclear import or retention of Drosha. Furthermore, depletion of Pasha or Drosha causes both components of the Microprocessor to mislocalize to the cytoplasm. Thus, Pasha and Drosha mutually regulate each other’s spatial expression inC. elegans.

Publisher

Cold Spring Harbor Laboratory

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