期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 293, 期 51, 页码 19522-19531出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.RA118.005120
关键词
nuclear magnetic resonance (NMR); intrinsically disordered protein; heterogeneous nuclear ribonucleoprotein (hnRNP); RNA transport; structural biology; protein-protein interaction; liquid-liquid phase separation (LLPS); membraneless organelle; prion-like domain; stress granule; aggregation; hnRNPA2; Fyn-SH3
资金
- NIGMS, National Institutes of Health [R01GM118530, P20GM104937]
- Karen T. Romer Undergraduate Teaching and Research Awards
- Robert J. and Nancy D. Carney Institute for Brain Science at Brown University [T32MH020068]
- NINDS, National Institutes of Health [F31NS110301]
Liquid-liquid phase separation of proteins and nucleic acids into membraneless organelles (MLOs) spatially organizes cellular components and reactions. The RNA-binding protein heterogeneous nuclear ribonucleoprotein A2 (hnRNPA2) carries mRNA targets in MLOs called transport granules in neurons and oligodendrocytes. At sites of local translation, hnRNPA2 is phosphorylated by the tyrosine protein kinase Fyn, releasing the mRNA for translation. Fyn recognizes targets through its SH3 domain (Fyn-SH3). However, hnRNPA2 lacks canonical SH3-binding sequences, raising the question of how Fyn-SH3 binds hnRNPA2 in phase-separated transport granules. Here, we characterize the structural details of the interaction of the hnRNPA2 low-complexity domain (LC) with Fyn-SH3 and the effect of Fyn-SH3 on hnRNPA2 phase separation. We combined in vitro microscopy and solution NMR spectroscopy to evaluate assembly of hnRNPA2 and Fyn-SH3 into in vitro phase-separated granules and probe the structural details of their interaction. We observed that Fyn-SH3 induces hnRNPA2 LC phase separation and that Fyn-SH3 is incorporated into in vitro hnRNPA2 LC granules. Moreover, we identified hnRNPA2 LC interaction sites on the surface of Fyn-SH3. Our data offer a structural view of how hnRNPA2 LC may interact with Fyn. To our knowledge, our study provides the first example of a single globular domain inducing phase separation of a disordered MLO scaffold protein.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据