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DOI10.1073/pnas.2015621118
Distinct roles of nuclear basket proteins in directing the passage of mRNA through the nuclear pore
Li Y.; Aksenova V.; Tingey M.; Yu J.; Ma P.; Arnaoutov A.; Chen S.; Dasso M.; Yang W.
Date Issued2021
ISSN0027-8424
Volume118Issue:37
Other AbstractThe in vivo characterization of the exact copy number and the specific function of each composite protein within the nuclear pore complex (NPC) remains both desirable and challenging. Through the implementation of live-cell high-speed super-resolution singlemolecule microscopy, we first quantified the native copies of nuclear basket (BSK) proteins (Nup153, Nup50, and Tpr) prior to knocking them down in a highly specific manner via an auxin-inducible degron strategy. Second, we determined the specific roles that BSK proteins play in the nuclear export kinetics ofmodelmessenger RNA (mRNA) substrates. Finally, the three-dimensional (3D) nuclear export routes of these mRNA substrates through native NPCs in the absence of specific BSK proteins were obtained and further validated via postlocalization computational simulations. We found that these BSK proteins possess the stoichiometric ratio of 1:1:1 and play distinct roles in the nuclear export of mRNAs within live cells. The absence of Tpr from the NPC predominantly reduces the probability of nuclearmRNAs entering the NPC for export. Complete depletion of Nup153 and Nup50 results in an mRNA nuclear export efficiency decrease of approximately four folds. mRNAs can gain their maximum successful export efficiency as the copy number of Nup153 increased from zero to only half the full complement natively within the NPC. Lastly, the absence of Tpr or Nup153 seems to alter the 3D export routes of mRNAs as they pass through the NPC. However, the removal of Nup50 alone has almost no impact upon mRNA export route and kinetics. © 2021 National Academy of Sciences. All rights reserved.
enkeywords3D super-resolution microscopy; NPC stoichiometry; Nucleocytoplasmic transport
Language英语
journalProceedings of the National Academy of Sciences of the United States of America
Document Type期刊论文
Identifierhttp://gcip.llas.ac.cn/handle/2XKMVOVA/238801
AffiliationDepartment of Biology, Temple University, Philadelphia, PA 19122, United States; Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, MD 20847, United States
Recommended Citation
GB/T 7714
Li Y.,Aksenova V.,Tingey M.,et al. Distinct roles of nuclear basket proteins in directing the passage of mRNA through the nuclear pore[J],2021,118(37).
APA Li Y..,Aksenova V..,Tingey M..,Yu J..,Ma P..,...&Yang W..(2021).Distinct roles of nuclear basket proteins in directing the passage of mRNA through the nuclear pore.Proceedings of the National Academy of Sciences of the United States of America,118(37).
MLA Li Y.,et al."Distinct roles of nuclear basket proteins in directing the passage of mRNA through the nuclear pore".Proceedings of the National Academy of Sciences of the United States of America 118.37(2021).
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