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Liquid droplet germ granules require assembly and localized regulators for mRNA repression

Scott Takeo Aoki, Tina R Lynch, Sarah L Crittenden, Craig A Bingman, Marvin Wickens, Judith Kimble
doi: https://doi.org/10.1101/382838
Scott Takeo Aoki
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
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Tina R Lynch
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
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Sarah L Crittenden
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
2Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, WI, USA
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Craig A Bingman
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
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Marvin Wickens
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
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Judith Kimble
1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA
2Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, WI, USA
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  • For correspondence: jekimble@wisc.edu
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Summary

Cytoplasmic RNA-protein (RNP) granules have diverse biophysical properties, from liquid to solid, and play enigmatic roles in RNA metabolism. Nematode P-granules are paradigmatic liquid droplet granules and central to germ cell development. Here we analyze a key P-granule scaffolding protein, called PGL, to investigate the functional relationship between P-granule assembly and function. Using a protein-RNA tethering assay, we find that reporter mRNA expression is repressed when recruited to PGL granules. We determine the crystal structure of the PGL N-terminal region to 1.5 Å, discover its dimerization and identify key residues at the dimer interface. In vivo mutations of those interface residues prevent P-granule assembly, de-repress PGL-tethered mRNA and reduce fertility. Therefore, PGL dimerization lies at the heart of both P-granule assembly and function. Finally, we identify the P-granule-associated Argonaute WAGO-1 as crucial for repression of PGL-tethered mRNA. We conclude that P-granule function requires both assembly and localized regulators.

Footnotes

  • Abbreviations: RNA, ribonucleic acid; UTR, untranslated region; GFP, green fluorescent protein; CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats; RNAi, ribonucleic acid interference

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted July 05, 2019.
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Liquid droplet germ granules require assembly and localized regulators for mRNA repression
Scott Takeo Aoki, Tina R Lynch, Sarah L Crittenden, Craig A Bingman, Marvin Wickens, Judith Kimble
bioRxiv 382838; doi: https://doi.org/10.1101/382838
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Liquid droplet germ granules require assembly and localized regulators for mRNA repression
Scott Takeo Aoki, Tina R Lynch, Sarah L Crittenden, Craig A Bingman, Marvin Wickens, Judith Kimble
bioRxiv 382838; doi: https://doi.org/10.1101/382838

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