Perelman Rebecca T, Schmidt Andreas, Khan Umar, Walter Nils G
Single Molecule Analysis Group, University of Michigan, Ann Arbor, MI 48109, USA.
Center for Advanced Biomedical Imaging and Photonics, Beth Israel Deaconess Medical Center, Harvard University, Boston, MA 02115, USA.
Cells. 2023 Sep 11;12(18):2250. doi: 10.3390/cells12182250.
Cellular biomolecular condensates, termed ribonucleoprotein (RNP) granules, are often enriched in messenger RNA (mRNA) molecules relative to the surrounding cytoplasm. Yet, the spatial localization and diffusion of mRNAs in close proximity to phase separated RNP granules are not well understood. In this study, we performed single-molecule fluorescence imaging experiments of mRNAs in live cells in the presence of two types of RNP granules, stress granules (SGs) and processing bodies (PBs), which are distinct in their molecular composition and function. We developed a photobleaching- and noise-corrected colocalization imaging algorithm that was employed to determine the accurate positions of individual mRNAs relative to the granule's boundaries. We found that mRNAs are often localized at granule boundaries, an observation consistent with recently published data. We suggest that mRNA molecules become spontaneously confined at the RNP granule boundary similar to the adsorption of polymer molecules at liquid-liquid interfaces, which is observed in various technological and biological processes. We also suggest that this confinement could be due to a combination of intermolecular interactions associated with, first, the screening of a portion of the RNP granule interface by the polymer and, second, electrostatic interactions due to a strong electric field induced by a Donnan potential generated across the thin interface.
被称为核糖核蛋白(RNP)颗粒的细胞生物分子凝聚物,相对于周围的细胞质,通常富含信使核糖核酸(mRNA)分子。然而,mRNA在相分离的RNP颗粒附近的空间定位和扩散情况尚未得到很好的理解。在本研究中,我们在存在两种RNP颗粒(应激颗粒(SGs)和加工小体(PBs))的情况下,对活细胞中的mRNA进行了单分子荧光成像实验,这两种颗粒在分子组成和功能上有所不同。我们开发了一种经过光漂白和噪声校正的共定位成像算法,用于确定单个mRNA相对于颗粒边界的准确位置。我们发现mRNA常常定位在颗粒边界,这一观察结果与最近发表的数据一致。我们认为,mRNA分子会自发地被限制在RNP颗粒边界,类似于聚合物分子在液 - 液界面的吸附,这种现象在各种技术和生物过程中都有观察到。我们还认为,这种限制可能是由于分子间相互作用的组合,首先是聚合物对RNP颗粒界面一部分的屏蔽,其次是由于在薄界面上产生的唐南电位诱导的强电场导致的静电相互作用。