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Article ; Online: In vitro characterization of the full-length human dynein-1 cargo adaptor BicD2.

Fagiewicz, Robert / Crucifix, Corinne / Klos, Torben / Deville, Célia / Kieffer, Bruno / Nominé, Yves / Busselez, Johan / Rossolillo, Paola / Schmidt, Helgo

Structure (London, England : 1993)

2022  Volume 30, Issue 11, Page(s) 1470–1478.e3

Abstract: Cargo adaptors are crucial in coupling motor proteins with their respective cargos and regulatory proteins. BicD2 is a prominent example within the cargo adaptor family. BicD2 is able to recruit the microtubule motor dynein to RNA, viral particles, and ... ...

Abstract Cargo adaptors are crucial in coupling motor proteins with their respective cargos and regulatory proteins. BicD2 is a prominent example within the cargo adaptor family. BicD2 is able to recruit the microtubule motor dynein to RNA, viral particles, and nuclei. The BicD2-mediated interaction between the nucleus and dynein is implicated in mitosis, interkinetic nuclear migration (INM) in radial glial progenitor cells, and neuron precursor migration during embryonic neocortex development. In vitro studies involving full-length cargo adaptors are difficult to perform due to the hydrophobic character, low-expression levels, and intrinsic flexibility of cargo adaptors. Here, we report the recombinant production of full-length human BicD2 and confirm its biochemical activity by interaction studies with RanBP2. We also describe pH-dependent conformational changes of BicD2 using cryoelectron microscopy (cryo-EM), template-free structure predictions, and biophysical tools. Our results will help define the biochemical parameters for the in vitro reconstitution of higher-order BicD2 protein complexes.
MeSH term(s) Humans ; Dyneins/metabolism ; Dynactin Complex/metabolism ; Microtubule-Associated Proteins/metabolism ; Cryoelectron Microscopy ; Microtubules/metabolism
Chemical Substances Dyneins (EC 3.6.4.2) ; Dynactin Complex ; Microtubule-Associated Proteins ; BICD2 protein, human
Language English
Publishing date 2022-09-22
Publishing country United States
Document type Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID 1213087-4
ISSN 1878-4186 ; 0969-2126
ISSN (online) 1878-4186
ISSN 0969-2126
DOI 10.1016/j.str.2022.08.009
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