The Multivalent Polyampholyte Domain of Nst1, a P-Body-Associated Saccharomyces cerevisiae Protein, Provides a Platform for Interacting with P-Body Components
Abstract
:1. Introduction
2. Results
2.1. The Nst1 C-Terminal Domain (CTD) Contains Polyampholyte and Aggregation-Prone Regions
2.2. The Nst1 CTD Is Sufficient for Nst1 Self-Condensation
2.3. The APD in the Nst1 CTD Is Insufficient but Crucial for Inducing Nst1 Self-Condensation
2.4. The Nst1 PD Is Not a Critical Component in Self-Condensation but Is Responsible for Inducing Dcp2 Condensation
2.5. Dcp2 Condensation Induced by Nst1 PD Overexpression Is Independent of Free Ribosomal Influx
2.6. The Nst1 PD Serves as a Binding Hub, Mediating the Condensation of other PB Components
3. Discussion
3.1. The Nst1 C-Terminus Is Necessary and Sufficient for Self-Condensation, While the N-Terminus Has an Auxiliary Role in Recruiting other PB Components
3.2. The Aggregation-Prone Region May Be Associated with Inducing Nst1 Condensates with Liquid-like Properties
3.3. The Polyampholyte Region May Be Involved in Molecular Condensation as a Platform for Multivalent Protein–Protein Interactions Independent of RNA Influx
4. Materials and Methods
4.1. Yeast strains, Plasmids, and Cultures
4.2. 1,6-Hexanediol and CHX Treatments and Western Blots
4.3. Nst1 Structure and Domain Predictions Based on the Sequence
4.4. Wide-Field Fluorescence Microscopy of Yeast Cells and Image Analysis
4.5. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strain Name | Genotype | Source |
---|---|---|
YSK3485 | DCP2-EGFP:HIS3MX6 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
YSK3482 | XRN1-EGFP::HIS3MX6 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
YSK3484 | DHH1-EGFP:HIS3MX6 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
YSK3534 | EDC3-EGFP::HIS3MX6 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
YSK3578 | DCP2-mKate2-sphis5 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
YSK3483 | BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 wild-type | This study |
YSK3592 | DCP2-9MYC::HIS3MX6 BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 | This study |
Plasmids |
---|
pMW20-PGAL-GFP-NST |
pMW20-PGAL-GFP-NSTΔ430–1240 |
pMW20-PGAL-GFP-NST1Δ1–429 |
pMW20-PGAL-GFP-NST1Δ1–429 Δ631–752 |
pMW20-PGAL-GFP-NST1Δ1–429 Δ1016–1240 |
pMW20-PGAL-GFP-NST1Δ1–1015 |
pMW20-PGAL-GFP-NST1Δ631–752 |
pMW20-PGAL-GFP-NST1Δ631–752 Δ1016–1240 |
pMW20-PGAL-GFP-NST1Δ1016–1240 |
pMW20-PGAL-GFP-NST1Δ1–1015 |
pMW20-PGAL-NST |
pMW20-PGAL-NSTΔ430–1240 |
pMW20-PGAL-NST1Δ1–429 |
pMW20-PGAL-NST1Δ1–429 Δ631–752 |
pMW20-PGAL-NST1Δ1–429 Δ1016–1240 |
pMW20-PGAL-NST1Δ631–752 |
pMW20-PGAL-NST1Δ631–752 Δ1016–1240 |
pMW20-PGAL-NST1Δ1016–1240 |
pMW20-PGAL-NST1Δ1–429 Δ631–752 |
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Choi, Y.-J.; Lee, Y.; Lin, Y.; Heo, Y.; Lee, Y.-H.; Song, K. The Multivalent Polyampholyte Domain of Nst1, a P-Body-Associated Saccharomyces cerevisiae Protein, Provides a Platform for Interacting with P-Body Components. Int. J. Mol. Sci. 2022, 23, 7380. https://doi.org/10.3390/ijms23137380
Choi Y-J, Lee Y, Lin Y, Heo Y, Lee Y-H, Song K. The Multivalent Polyampholyte Domain of Nst1, a P-Body-Associated Saccharomyces cerevisiae Protein, Provides a Platform for Interacting with P-Body Components. International Journal of Molecular Sciences. 2022; 23(13):7380. https://doi.org/10.3390/ijms23137380
Chicago/Turabian StyleChoi, Yoon-Jeong, Yujin Lee, Yuxi Lin, Yunseok Heo, Young-Ho Lee, and Kiwon Song. 2022. "The Multivalent Polyampholyte Domain of Nst1, a P-Body-Associated Saccharomyces cerevisiae Protein, Provides a Platform for Interacting with P-Body Components" International Journal of Molecular Sciences 23, no. 13: 7380. https://doi.org/10.3390/ijms23137380