title: Coexisting Liquid Phases Underlie Nucleolar Subcompartments.
abstract: The nucleolus and other ribonucleoprotein (RNP) bodies are membrane-less organelles that appear to assemble through phase separation of their molecular components. However, many such RNP bodies contain internal subcompartments, and the mechanism of their formation remains unclear. Here, we combine in vivo and in vitro studies, together with computational modeling, to show that subcompartments within the nucleolus represent distinct, coexisting liquid phases. Consistent with their in vivo immiscibility, purified nucleolar proteins phase separate into droplets containing distinct non-coalescing phases that are remarkably similar to nucleoli in vivo. This layered droplet organization is caused by differences in the biophysical properties of the phases-particularly droplet surface tension-which arises from sequence-encoded features of their macromolecular components. These results suggest that phase separation can give rise to multilayered liquids that may facilitate sequential RNA processing reactions in a variety of RNP bodies. PAPERCLIP.
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Feric
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
authors.forename: Nilesh
authors.initials: N
authors.lastname: Vaidya
authors.affiliation: Department of Physics, Washington University in St. Louis, St. Louis, MO 63130, USA; Department of Biomedical Engineering and Center for Biological Systems Engineering, Washington University in St. Louis, St. Louis, MO 63130, USA.
authors.forename: Tyler S
authors.initials: TS
authors.lastname: Harmon
authors.affiliation: Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38103, USA.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Mitrea
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
authors.forename: Lian
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
authors.forename: Tiffany M
authors.initials: TM
authors.lastname: Richardson
authors.affiliation: Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38103, USA.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Kriwacki
authors.affiliation: Department of Biomedical Engineering and Center for Biological Systems Engineering, Washington University in St. Louis, St. Louis, MO 63130, USA.
authors.forename: Rohit V
authors.initials: RV
authors.lastname: Pappu
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA. Electronic address: cbrangwy@princeton.edu.
authors.forename: Clifford P
authors.initials: CP
authors.lastname: Brangwynne
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans; D002466:Cell Nucleolus / Q000737:chemistry*; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis; D007422:Intestines / Q000737:chemistry / Q000166:cytology; D008322:Mammals; D009687:Nuclear Proteins / Q000032:analysis; D000090243:Nucleophosmin; D009865:Oocytes / Q000737:chemistry / Q000166:cytology; D012323:RNA Processing, Post-Transcriptional; D012261:Ribonucleoproteins / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; C047458:fibrillarin; D000090243:Nucleophosmin