Cell cycle-specific phase separation regulated by protein charge blockiness

May 13, 2022

Title

Cell cycle-specific phase separation regulated by protein charge blockiness

Author

Hiroya Yamazaki, Masatoshi Takagi, Hidetaka Kosako, Tatsuya Hirano & Shige H. Yoshimura

Year

2022

Journal

Nature Cell Biology

Abstract

Dynamic morphological changes of intracellular organelles are often regulated by protein phosphorylation or dephosphorylation1,2,3,4,5,6. Phosphorylation modulates stereospecific interactions among structured proteins, but how it controls molecular interactions among unstructured proteins and regulates their macroscopic behaviours remains unknown. Here we determined the cell cycle-specific behaviour of Ki-67, which localizes to the nucleoli during interphase and relocates to the chromosome periphery during mitosis. Mitotic hyperphosphorylation of disordered repeat domains of Ki-67 generates alternating charge blocks in these domains and increases their propensity for liquid–liquid phase separation (LLPS). A phosphomimetic sequence and the sequences with enhanced charge blockiness underwent strong LLPS in vitro and induced chromosome periphery formation in vivo. Conversely, mitotic hyperphosphorylation of NPM1 diminished a charge block and suppressed LLPS, resulting in nucleolar dissolution. Cell cycle-specific phase separation can be modulated via phosphorylation by enhancing or reducing the charge blockiness of disordered regions, rather than by attaching phosphate groups to specific sites.

Instrument

V-630

Keywords

protein, structure, integration, LLPS