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Analysis of Spliceosomal snRNA Localization in Human Hela Cells Using Microinjection
Published on: August 6, 2019
The Sam68 nuclear body is composed of two RNase-sensitive substructures joined by the adaptor HNRNPL
Taro Mannen1, Seisuke Yamashita2, Kozo Tomita2
1Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
Abstract:
The mammalian cell nucleus contains membraneless suborganelles referred to as nuclear bodies (NBs). Some NBs are formed with an architectural RNA (arcRNA) as the structural core. Here, we searched for new NBs that are built on unidentified arcRNAs by screening for ribonuclease (RNase)-sensitive NBs using 32,651 fluorescently tagged human cDNA clones. We identified 32 tagged proteins that required RNA for their localization in distinct nuclear foci. Among them, seven RNA-binding proteins commonly localized in the Sam68 nuclear body (SNB), which was disrupted by RNase treatment. Knockdown of each SNB protein revealed that SNBs are composed of two distinct RNase-sensitive substructures. One substructure is present as a distinct NB, termed the DBC1 body, in certain conditions, and the more dynamic substructure including Sam68 joins to form the intact SNB. HNRNPL acts as the adaptor to combine the two substructures and form the intact SNB through the interaction of two sets of RNA recognition motifs with the putative arcRNAs in the respective substructures.
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