Details

Autor(en) / Beteiligte

• Drew, Kevin
• Lee, Chanjae
• Cox, Rachael M.
• Dang, Vy
• Devitt, Caitlin C.
• McWhite, Claire D.
• Papoulas, Ophelia
• Huizar, Ryan L.
• Marcotte, Edward M.
• Wallingford, John B.

Titel

A systematic, label-free method for identifying RNA-associated proteins in vivo provides insights into vertebrate ciliary beating machinery

Ist Teil von

• Developmental biology, 2020-11, Vol.467 (1-2), p.108-117

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2020

Link zum Volltext

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• Cell-type specific RNA-associated proteins are essential for development and homeostasis in animals. Despite a massive recent effort to systematically identify RNA-associated proteins, we currently have few comprehensive rosters of cell-type specific RNA-associated proteins in vertebrate tissues. Here, we demonstrate the feasibility of determining the RNA-associated proteome of a defined vertebrate embryonic tissue using DIF-FRAC, a systematic and universal (i.e., label-free) method. Application of DIF-FRAC to cultured tissue explants of Xenopus mucociliary epithelium identified dozens of known RNA-associated proteins as expected, but also several novel RNA-associated proteins, including proteins related to assembly of the mitotic spindle and regulation of ciliary beating. In particular, we show that the inner dynein arm tether Cfap44 is an RNA-associated protein that localizes not only to axonemes, but also to liquid-like organelles in the cytoplasm called DynAPs. This result led us to discover that DynAPs are generally enriched for RNA. Together, these data provide a useful resource for a deeper understanding of mucociliary epithelia and demonstrate that DIF-FRAC will be broadly applicable for systematic identification of RNA-associated proteins from embryonic tissues. •Demonstrates the feasibility of determining the RNA-associated proteome of a defined vertebrate embryonic tissue.•Identified novel RNA-associated proteins related to assembly of the mitotic spindle and regulation of ciliary beating.•Cfap44 is an RNA-associated protein that localizes to RNA-based liquid-like organelles in the cytoplasm called DynAPs.