Recruitment of Two Dyneins to an mRNA-Dependent Bicaudal D Transport Complex

Sladewski, Thomas E.; Billington, Neil; Ali, M. Yusuf; Bookwalter, Carol S.; Lu, Hailong; Krementsova, Elena B.; Schroer, Trina A.; Trybus, Kathleen M.

doi:10.1101/273755

Cited by 24 publications

(85 citation statements)

References 52 publications

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“…In the absence of cargo, BicD forms an autoinhibited state that is unable to recruit dynein (3)(4)(5)(6)(7)(8)(9)(10)(11)(12). Here we show by single molecule processivity assays that auto-inhibition is abolished in the classical Bicaudal mutant BicD/F684I, resulting in cargo-independent activation of dynein-dynactin, consistent with cellular studies that show increased dynein recruitment (9,13).…”

Section: Discussionsupporting

confidence: 87%

“…Dynein was expressed with an N-terminal biotin tag in Sf9 cells and dynactin was purified from bovine brain tissue as described (11). Fulllength Drosophila BicD WT (BicD WT ) and BicD/F684I (BicD F684I ) were expressed in Sf9 cells and BicD-coiled-coil domain 1 (BicD CC1 ) in bacteria as described (11).…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

“…The motility assay was performed on PEGylated slides using a buffer containing 30 mM HEPES pH 7.4, 50 mM KOAc, 2 mM MgOAc, 1 mM EGTA, 2 mM MgATP, 20 mM DTT, 8 mg/ml BSA, 0.5 mg/ml k-casein, 0.5% pluronic F68, 10 mM paclitaxel and an oxygen scavenger system (5.8 mg/ml glucose, 0.045 mg/ml catalase, and 0.067 mg/ml glucose oxidase (11). Total Internal Reflectance Fluorescence (TIRF) microscopy was used to capture images of Qdot labeled dynein.…”

Section: Single Molecule Processivity Assays With Fulllength Dm Bicdmentioning

confidence: 99%

“…The CTD is required for autoinhibition, since a truncated BicD-CC1 construct without it activates dynein for processive transport in the absence of cargo (5,7). Auto-inhibition of full-length BicD is released upon cargo binding (3)(4)(5)(6)(7)(8)(9)(10)(11)(12); however, the underlying molecular mechanism is elusive.…”

mentioning

confidence: 99%

“…Cargo-bound adaptors are required to activate metazoan dynein for processive transport and are therefore an essential part of the dynein transport machinery (3)(4)(5)(6)(7)(8). In the absence of cargo, BicD forms an auto-inhibited looped conformation, in which the C-terminal cargo binding region (CTD) binds to the Nterminal dynein/dynactin-binding site (coiled-coil domain 1, CC1), sterically preventing motor binding (3)(4)(5)(6)(7)(8)(9)(10)(11)(12). The CTD is required for autoinhibition, since a truncated BicD-CC1 construct without it activates dynein for processive transport in the absence of cargo (5,7).…”

mentioning

confidence: 99%

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Coiled-coil registry shifts in the F684I mutant of Bicaudal result in cargo-independent activation of dynein motility

Cui

Trybus

Ali

et al. 2019

Preprint

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ABSTRACTThe dynein adaptor Drosophila Bicaudal D (BicD) is auto-inhibited and activates dynein motility only after cargo is bound, but the underlying mechanism is elusive. In contrast, we show that the full-length BicD/F684I mutant activates dynein processivity even in the absence of cargo. Our X-ray structure of the C-terminal domain of the BicD/F684I mutant reveals a coiled-coil registry shift; in the N-terminal region, the two helices of the homodimer are aligned, whereas they are vertically shifted in the wild-type. One chain is partially disordered and this structural flexibility is confirmed by computations, which reveal that the mutant transitions back and forth between the two registries. We propose that a coiled-coil registry shift upon cargo binding activates BicD for dynein recruitment. Moreover, the human homolog BicD2/F743I exhibits diminished binding of cargo adaptor Nup358, implying that a coiled-coil registry shift may be a mechanism to modulate cargo selection for BicD2–dependent transport pathways.

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Section: Discussionsupporting

confidence: 87%

Section: Protein Expression and Purificationmentioning

confidence: 99%

Section: Single Molecule Processivity Assays With Fulllength Dm Bicdmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Coiled-coil registry shifts in the F684I mutant of Bicaudal result in cargo-independent activation of dynein motility

Cui

Trybus

Ali

et al. 2019

Preprint

Self Cite

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Patterning the Drosophila embryo: A paradigm for RNA‐based developmental genetic regulation

Lasko

2020

WIREs RNA

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Embryonic anterior-posterior patterning is established in Drosophila melanogaster by maternally expressed genes. The mRNAs of several of these genes accumulate at either the anterior or posterior pole of the oocyte via a number of mechanisms. Many of these mRNAs are also under elaborate translational regulation. Asymmetric RNA localization coupled with spatially restricted translation ensures that their proteins are restricted to the position necessary for the developmental process that they drive. Bicoid (Bcd), the anterior determinant, and Oskar (Osk), the determinant for primordial germ cells and posterior patterning, have been studied particularly closely. In early embryos an anterior-posterior gradient of Bcd is established, activating transcription of different sets of zygotic genes depending on local Bcd concentration. At the posterior pole, Osk seeds formation of polar granules, ribonucleoprotein complexes that accumulate further mRNAs and proteins involved in posterior patterning and germ cell specification. After fertilization, polar granules associate with posterior nuclei and mature into nuclear germ granules. Osk accumulates in these granules, and either by itself or as part of the granules, stimulates germ cell division.

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