CRACR2a is a calcium-activated dynein adaptor protein that regulates endocytic traffic

Wang, Yuxiao; Huynh, Walter; Skokan, Taylor D.; Lü, Wen; Weiss, Arthur; Vale, Ronald D.

doi:10.1083/jcb.201806097

Cited by 52 publications

(85 citation statements)

References 44 publications

(89 reference statements)

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“…We hypothesize that related activating adaptors or candidate activating adaptors will also require Lis1 to maximally activate dynein/ dynactin. In humans there are three additional BicD family members (BicD1, BicDL1, and BicDL2), two additional Hook family members (Hook1 and Hook2), and additional proteins that share a similar domain structure in their dynein/ dynactin binding regions to the Hook proteins (CCDC88A, CCDC88B, and CCDC88C) or Ninl (Nin, Rab11-FIP3, CRACR2a, Rab 44, and Rab 45) 2,9,52 . Thus, we predict that Lis1 will play a role in the cell biological processes that all of these activating adaptors or candidate activating adaptors facilitate.…”

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confidence: 99%

Lis1 promotes the formation of maximally activated cytoplasmic dynein-1 complexes

Htet

Gillies

Baker

et al. 2019

Preprint

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Cytoplasmic dynein-1 is a molecular motor that drives nearly all minus-end-directed microtubule-based transport in human cells, performing functions ranging from retrograde axonal transport to mitotic spindle assembly 1,2 . Activated dynein complexes consist of one or two dynein dimers, the dynactin complex, and an "activating adaptor", with maximal velocity seen with two dimers present ( Fig. 1a) 3-6 . Little is known about how this massive ~4MDa complex is assembled. Using purified recombinant human proteins, we uncovered a novel role for the dynein-binding protein, Lis1, in the formation of fully activated dynein complexes containing two dynein dimers. Lis1 is required for maximal velocity of complexes activated by proteins representing three different families of activating adaptors: BicD2, Hook3, and Ninl. Once activated dynein complexes have formed, they do not require the presence of Lis1 for sustained maximal velocity. Using cryo-electron microscopy we show that human Lis1 binds to dynein at two sites on dynein's motor domain, similar to yeast dynein 7 . We propose that the ability of Lis1 to bind at these sites may function in multiple stages of assembling the motile human dynein/ dynactin/ activating adaptor complex.

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confidence: 99%

Lis1 promotes the formation of maximally activated cytoplasmic dynein-1 complexes

Htet

Gillies

Baker

et al. 2019

Preprint

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“…Our previous findings revealed a dynein-dependent retrograde trafficking of WPBs to the MTOC that required Rab46 GTPase activity (Miteva et al, 2019). Moreover, the identification of Rab46 as a new dynein adaptor protein in immune cells by Wang et al, (Wang et al, 2019) advocates DHC as a promising candidate for further investigation.…”

Section: Rab46 Interacts Directly With the Dynein Complexmentioning

confidence: 86%

“…Rab46 contains two functional Ca 2+ binding motifs (EF-hands) in the N-terminal (Srikanth et al, 2010). Wang et al recently reported that Ca 2+ binding to the EF-hands was necessary for the interaction between Rab46 and dynein in T-cells (Wang et al, 2019). Surprisingly, in ECs, chelation of intracellular Ca 2+ does not inhibit dynein-dependent trafficking of WPBs to the MTOC (Miteva et al, 2019).…”

Section: Rab46 / Dynein Interaction Is Independent Of Calciummentioning

confidence: 99%

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Identification of novel Rab46 effector proteins

Pedicini

Wiktor

Simmons

et al. 2020

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AbstractRab46 is a novel Ca2+- sensing Rab GTPase shown to have important functions in endothelial and immune cells. The presence of functional Ca2+- binding, coiled-coil and Rab domains suggest that Rab46 will be important for coupling rapid responses to signalling in many cell types. The molecular mechanisms underlying Rab46 function are currently unknown. Here we provide the first resource for studying Rab46 interacting proteins. Using liquid chromatography mass spectrometry (LC-MS/MS) to identify affinity purified proteins that bind to constitutively active GFP-Rab46 or inactive GFP-Rab46 expressed in endothelial cells, we have revealed 922 possible interacting proteins. Further comparative analyses between nucleotide-locked Rab46 proteins enriched this dataset to confidently identify 29 effector proteins. Importantly, through biochemical and imaging approaches we have validated two potential effector proteins; dynein and the Na2+/ K+ ATPase subunit alpha 1 (ATP1α1). Hence, our use of affinity purification and LC-MS/MS to identify Rab46 neighbouring proteins provides a valuable resource for detecting Rab46 effector proteins and analysing Rab46 functions.

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“…Cytoplasmic dynein complexes are composed of dynein intermediate chains, light-intermediate chains (LIC) and light chains, which form a complex with two identical subunits of dynein heavy chains 16 . The complex is regulated by dynactin and adaptor proteins [17][18][19] , which facilitate the movement along microtubules 20 , but how dynein light chain subunits help perform individual cellular functions is not completely understood.…”

Section: Introductionmentioning

confidence: 99%

DYNC1LI2 regulates localization of the chaperone-mediated autophagy-receptor LAMP2A and improves cellular homeostasis in cystinosis

Rahman

Johnson

Zhang

et al. 2020

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The dynein motor protein complex is required for retrograde transport but the functions of the intermediate-light chains that form the cargo-binding complex are not elucidated and the importance of individual subunits in the maintenance of cellular homeostasis is unknown. Here, using mRNA arrays and protein analysis, we show that the dynein subunit, intermediate chain 2 (DYNC1LI2) is downregulated in cystinosis, a lysosomal storage disorder caused by genetic defects in the lysosomal cystine transporter, cystinosin. Reconstitution of the expression of DYNC1LI2 in Ctns -/cells re-established endolysosomal dynamics. Defective vesicular trafficking in cystinotic cells was rescued by DYNC1LI2 expression which correlated with decreased endoplasmic reticulum stress manifested as decreased expression levels of the chaperone Grp78.Mitochondrial fragmentation in cystinotic fibroblasts was also rescued by DYNC1LI2. Survival of cystinotic cells to oxidative stress insult was increased by DYNC1LI2 reconstitution but not by its paralog DYNC1LI1, which also failed to decrease ER stress levels and mitochondrial fragmentation. Restoring DYNC1LI2 expression rescued the localization of the chaperonemediated autophagy receptor, LAMP2A, and restored cellular homeostasis of cystinotic proximal tubule cells, the primary cell type affected in cystinosis. DYNC1LI2 failed to rescue phenotypes in cystinotic cells when LAMP2A was downregulated or when co-expressed with dominant negative (DN) RAB7 or DN-RAB11, which impair LAMP2A trafficking. DYNC1LI2 emerges as a new target to repair underlying trafficking and CMA defects in cystinosis, a mechanism that is not restored by currently used lysosomal depletion therapies.2

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CRACR2a is a calcium-activated dynein adaptor protein that regulates endocytic traffic

Cited by 52 publications

References 44 publications

Lis1 promotes the formation of maximally activated cytoplasmic dynein-1 complexes

Lis1 promotes the formation of maximally activated cytoplasmic dynein-1 complexes

Identification of novel Rab46 effector proteins

DYNC1LI2 regulates localization of the chaperone-mediated autophagy-receptor LAMP2A and improves cellular homeostasis in cystinosis

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