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Intrinsically disordered proteins SAID1/2 condensate on SERRATE/ARS2 for dual inhibition of miRNA biogenesis in Arabidopsis

Shang, B.; Wang, L.; Yan, X.; Li, Y.; Li, C.; Wu, C.; Wang, T.; Guo, X.; Choi, S.; Zhang, T.; Wang, Z.; Tong, C.-Y.; Oh, T.; Zhang, X.; Wang, Z.; Peng, X.; Zhang, X.

2022-09-16 plant biology
10.1101/2022.09.13.507836 bioRxiv
Show abstract

Intrinsically disordered proteins (IDPs) SAID1/2 are hypothetic dentin sialophosphoprotein-like proteins, but their true functions are unknown. Here, we identified SAID1/2 as negative regulators of SERRATE(SE)/ARS2, a core factor in miRNA biogenesis complex (microprocessor). Loss-of-function double mutants of said1; said2 caused pleiotropic developmental defects and thousands of differentially-expressed genes that partially overlapped with those in se. said1; said2 also displayed increased assembly of microprocessor and elevated accumulation of miRNAs. Mechanistically, SAID1/2 promotes PRP4KA-mediated phosphorylation of SE, leading to its degradation in vivo. Unexpectedly, SAID1/2 have strong binding affinity to hairpin-structured pri-miRNAs and can sequester them from SE. Moreover, SAID1/2 directly inhibit pri-miRNA processing by microprocessor in vitro. Whereas SAID1/2 did not impact SE subcellular compartmentation, the proteins themselves exhibited liquid-liquid phase condensation that is nucleated on SE. Thus, we proposed that SAID1/2 reduce miRNA production through hijacking pri-miRNAs to prevent microprocessor activity while promoting SE phosphorylation and its destabilization in Arabidopsis.

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