PMID- 36972455
OWN - NLM
STAT- MEDLINE
DCOM- 20230329
LR  - 20230412
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 120
IP  - 14
DP  - 2023 Apr 4
TI  - Quantitative reconstitution of yeast RNA processing bodies.
PG  - e2214064120
LID - 10.1073/pnas.2214064120 [doi]
LID - e2214064120
AB  - Many biomolecular condensates appear to form through liquid-liquid phase 
      separation (LLPS). Individual condensate components can often undergo LLPS 
      in vitro, capturing some features of the native structures. However, natural 
      condensates contain dozens of components with different concentrations, dynamics, 
      and contributions to compartment formation. Most biochemical reconstitutions of 
      condensates have not benefited from quantitative knowledge of these cellular 
      features nor attempted to capture natural complexity. Here, we build on prior 
      quantitative cellular studies to reconstitute yeast RNA processing bodies (P 
      bodies) from purified components. Individually, five of the seven highly 
      concentrated P-body proteins form homotypic condensates at cellular protein and 
      salt concentrations, using both structured domains and intrinsically disordered 
      regions. Combining the seven proteins together at their cellular concentrations 
      with RNA yields phase-separated droplets with partition coefficients and dynamics 
      of most proteins in reasonable agreement with cellular values. RNA delays the 
      maturation of proteins within and promotes the reversibility of, P bodies. Our 
      ability to quantitatively recapitulate the composition and dynamics of a 
      condensate from its most concentrated components suggests that simple 
      interactions between these components carry much of the information that defines 
      the physical properties of the cellular structure.
FAU - Currie, Simon L
AU  - Currie SL
AUID- ORCID: 0000-0001-5083-1484
AD  - HHMI, UT Southwestern Medical Center, Dallas TX 75390.
AD  - Department of Biophysics, UT Southwestern Medical Center, Dallas, TX 75390.
FAU - Xing, Wenmin
AU  - Xing W
AUID- ORCID: 0000-0001-6445-0615
AD  - HHMI, UT Southwestern Medical Center, Dallas TX 75390.
AD  - Department of Biophysics, UT Southwestern Medical Center, Dallas, TX 75390.
FAU - Muhlrad, Denise
AU  - Muhlrad D
AD  - HHMI, University of Colorado, Boulder, CO 80309.
AD  - Department of Biochemistry, University of Colorado, Boulder, CO 80309.
FAU - Decker, Carolyn J
AU  - Decker CJ
AUID- ORCID: 0000-0002-2672-8919
AD  - HHMI, University of Colorado, Boulder, CO 80309.
AD  - Department of Biochemistry, University of Colorado, Boulder, CO 80309.
FAU - Parker, Roy
AU  - Parker R
AUID- ORCID: 0000-0002-8412-4152
AD  - HHMI, University of Colorado, Boulder, CO 80309.
AD  - Department of Biochemistry, University of Colorado, Boulder, CO 80309.
FAU - Rosen, Michael K
AU  - Rosen MK
AUID- ORCID: 0000-0002-0775-7917
AD  - HHMI, UT Southwestern Medical Center, Dallas TX 75390.
AD  - Department of Biophysics, UT Southwestern Medical Center, Dallas, TX 75390.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230327
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - *Processing Bodies
MH  - *Saccharomyces cerevisiae/genetics
MH  - RNA/genetics
PMC - PMC10083542
OTO - NOTNLM
OT  - P bodies
OT  - RNA
OT  - biomolecular condensates
OT  - phase separation
OT  - protein
COIS- The authors declare no competing interest.
EDAT- 2023/03/28 06:00
MHDA- 2023/03/29 06:05
PMCR- 2023/03/27
CRDT- 2023/03/27 15:33
PHST- 2023/03/29 06:05 [medline]
PHST- 2023/03/27 15:33 [entrez]
PHST- 2023/03/28 06:00 [pubmed]
PHST- 2023/03/27 00:00 [pmc-release]
AID - 202214064 [pii]
AID - 10.1073/pnas.2214064120 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2023 Apr 4;120(14):e2214064120. doi: 
      10.1073/pnas.2214064120. Epub 2023 Mar 27.