人源HP1α介导的相分离液滴形成的可视化与定量分析
Visualization and Quantitation of Phase-Separated Droplet Formation by Human HP1α.
作者信息
Keenen Madeline M, Larson Adam G, Narlikar Geeta J
机构信息
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, United States; Tetrad Graduate Program, University of California, San Francisco, San Francisco, CA, United States.
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, United States.
出版信息
Methods Enzymol. 2018;611:51-66. doi: 10.1016/bs.mie.2018.09.034. Epub 2018 Oct 31.
Abstract
The ability of the heterochromatin protein-1 (HP1) to phase separate into droplets suggests new mechanisms of gene organization in the cell nucleus. An accumulating body of work suggests that other nuclear proteins also display phase separation behaviors in vitro. To understand the mechanistic and biological significance of such droplet formation a rigorous biophysical characterization of this behavior is necessary. Herein we describe procedures for imaging HP1 droplets by brightfield microscopy, and two methods to quantify phase separation.
摘要
异染色质蛋白1（HP1）相分离形成液滴的能力揭示了细胞核中基因组织的新机制。越来越多的研究表明，其他核蛋白在体外也表现出相分离行为。为了理解这种液滴形成的机制和生物学意义，对这种行为进行严格的生物物理表征是必要的。在此，我们描述了通过明场显微镜对HP1液滴进行成像的方法，以及两种量化相分离的方法。
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