A microscopy-based screen employing multiplex genome sequencing identifies cargo-specific requirements for dynein velocity

Tan, Kaeling; Roberts, A. J.; Chonofsky, Mark; Egan, Martin J.; Reck-Peterson, Samara L.

doi:10.1091/mbc.e13-09-0557

Cited by 27 publications

(52 citation statements)

References 58 publications

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“…Cytoplasmic dynein and the plus-end-directed kinesin-3 drive bi-directional movements of early endosomes in fungi and metazoan [6, 34–36], and the role of kinesin-3 in early endosome transport was first discovered in U. maydis [35]. In U. maydis and A. nidulans , kinesin-3 drives early endosome movements toward the hyphal tip, whereas dynein drives them away from the hyphal tip, and therefore, an abnormal accumulation of early endosomes at the hyphal tip can be seen in mutants defective in dynein function [25, 26, 37–41]. Kinesin-3 dependent early endosome movement has also been found in Neurospora crassa [42].…”

Section: Cytoplasmic Dynein and Kinesin-3 Drive Bidirectional Movemenmentioning

confidence: 99%

“…The mechanism of nuclear distribution in filamentous fungi is not fully understood but appears to involve the role of dynein in regulating the dynamics of microtubules [24, 75, 78–81]. Interestingly, a recent screen for organelle distribution mutants in A. nidulans has identified two dynein HC mutations, in AAA1 and AAA3 respectively, which are more detrimental to early endosome migration than to nuclear migration [41]. Since analogous mutations in budding yeast dynein HC cause a significant reduction in the speed of dynein movement, these results indicate that a normal level of dynein motor activity is more crucial for early endosome movement than for nuclear migration [41].…”

Section: Mutations In the Motor And Tail Domains Of Dynein Heavy Chaimentioning

confidence: 99%

“…Interestingly, a recent screen for organelle distribution mutants in A. nidulans has identified two dynein HC mutations, in AAA1 and AAA3 respectively, which are more detrimental to early endosome migration than to nuclear migration [41]. Since analogous mutations in budding yeast dynein HC cause a significant reduction in the speed of dynein movement, these results indicate that a normal level of dynein motor activity is more crucial for early endosome movement than for nuclear migration [41]. In a different screen, a HC tail mutation was found to be important for both early endosome movement and nuclear distribution but did not seem to affect dynein complex assembly or dynein-dynactin interaction [40].…”

Section: Mutations In the Motor And Tail Domains Of Dynein Heavy Chaimentioning

confidence: 99%

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Cytoplasmic dynein and early endosome transport

Xiang

Qiu

Yao

et al. 2015

Cell. Mol. Life Sci.

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Microtubule-based distribution of organelles/vesicles is crucial for the function of many types of eukaryotic cells and the molecular motor cytoplasmic dynein is required for transporting a variety of cellular cargos toward the microtubule minus ends. Early endosomes represent a major cargo of dynein in filamentous fungi, and dynein regulators such as LIS1 and the dynactin complex are both required for early endosome movement. In fungal hyphae, kinesin-3 and dynein drive bi-directional movements of early endosomes. Dynein accumulates at microtubule plus ends; this accumulation depends on kinesin-1 and dynactin, and it is important for early endosome movements towards the microtubule minus ends. The physical interaction between dynein and early endosome requires the dynactin complex, and in particular, its p25 component. The FTS-Hook-FHIP (FHF) complex links dynein-dynactin to early endosomes, and within the FHF complex, Hook interacts with dynein-dynactin, and Hook-early endosome interaction depends on FHIP and FTS.

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Section: Cytoplasmic Dynein and Kinesin-3 Drive Bidirectional Movemenmentioning

confidence: 99%

Section: Mutations In the Motor And Tail Domains Of Dynein Heavy Chaimentioning

confidence: 99%

Section: Mutations In the Motor And Tail Domains Of Dynein Heavy Chaimentioning

confidence: 99%

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Cytoplasmic dynein and early endosome transport

Xiang

Qiu

Yao

et al. 2015

Cell. Mol. Life Sci.

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“…In a previous screen to identify novel regulators of microtubule-based transport, we mutagenized an A. nidulans strain expressing three fluorescently-labeled organelles known to be distributed by dynein and kinesin: EEs (GFP-RabA/5a), peroxisomes (peroxisome targeting signal mCherry-PTS1), and nuclei (Histone H1, HH1-BFP) [8]. From this screen, we identified two mutants with peroxisome-specific defects that mapped to two independent alleles in the gene AN1156/pxdA, which led to early stop codons (PxdAS201stop and PxdAQ846stop) in the protein ( Figure 1A) [39].…”

Section: Identification Of Novel Pxda Allelesmentioning

confidence: 99%

“…Methods for live-cell imaging in A. nidulans are wellestablished and the microtubules near the hyphal tip are uniformly polarized with their plus ends oriented outward, making the directionality of cargo transport and the motors involved easy to identify. We and others have exploited these features by performing forward genetic screens to identify regulators of microtubule-based transport [7][8][9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

PxdA interacts with the DipA phosphatase to regulate endosomal hitchhiking of peroxisomes

Salogiannis

Christensen

Aguilar-Maldonado

et al. 2020

Preprint

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The canonical mechanism of microtubule-based movement uses adaptor proteins to link cargos to the molecular motors dynein and kinesin. Recently, an alternative mechanism known as "hitchhiking" was discovered: ribonucleoproteins, peroxisomes, lipid droplets, and endoplasmic reticulum achieve motility by hitching a ride on motile early endosomes, rather than attaching directly to a motor protein. In the filamentous fungus Aspergillus nidulans we identified a molecular linker, PxdA, that associates with early endosomes and is essential for peroxisome hitchhiking. However, the molecular components that interact with PxdA, as well as the regulatory mechanisms that govern hitchhiking, are not understood. Here, we report the identification of two new pxdA alleles, including a point mutation (R2044P). We find that the R2044P mutation disrupts PxdA's ability to associate with early endosomes and consequently reduces peroxisome movement. We also identify the phosphatase DipA as an interaction partner of PxdA. DipA associates with early endosomes in a PxdA-dependent manner, and regulates the movement and distribution of peroxisomes. Finally, we find that PxdA also regulates the distribution of lipid droplets, but not autophagosomes or mitochondria. Our data suggest that PxdA is a central regulator of early endosome-dependent hitchhiking and requires the DipA phosphatase to regulate the movement and distribution of lipid droplets and peroxisomes.

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Activation of the motor protein upon attachment: Anchors weigh in on cytoplasmic dynein regulation

Parton

2016

BioEssays

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Cytoplasmic dynein is the major minus-end-directed motor protein in eukaryotes, and has functions ranging from organelle and vesicle transport to spindle positioning and orientation. The mode of regulation of dynein in the cell remains elusive, but a tantalising possibility is that dynein is maintained in an inhibited, non-motile state until bound to cargo. In vivo, stable attachment of dynein to the cell membrane via anchor proteins enables dynein to produce force by pulling on microtubules and serves to organise the nuclear material. Anchor proteins of dynein assume diverse structures and functions and differ in their interaction with the membrane. In yeast, the anchor protein has come to the fore as one of the key mediators of dynein activity. In other systems, much is yet to be discovered about the anchors, but future work in this area will prove invaluable in understanding dynein regulation in the cell.

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A microscopy-based screen employing multiplex genome sequencing identifies cargo-specific requirements for dynein velocity

Abstract: Methods are given for a screen using whole-genome multiplex sequencing in the filamentous fungus Aspergillus nidulans. Novel alleles of the molecular motor cytoplasmic dynein identified in this screen reveal that different dynein cargoes have distinct requirements for motor performance.

Cited by 27 publications

References 58 publications

Cytoplasmic dynein and early endosome transport

Cytoplasmic dynein and early endosome transport

PxdA interacts with the DipA phosphatase to regulate endosomal hitchhiking of peroxisomes

Activation of the motor protein upon attachment: Anchors weigh in on cytoplasmic dynein regulation

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