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In 2017 we reported a cryo-EM structure of full length cytoplasmic dynein in an inhibited state called the phi-particle. The two dynein heavy chains bind to accessory chains and wrap around each other to form the tail region before joining into the motor domains. Our structure showed how the packing of the motor domains locks them into a state that is unable to bind microtubules.<\/p>\n\n\n\n
In a collaboration with Ahmet Yildiz\u2019s lab we addressed the basis of dynein\u2019s directionality. We mutated the dynein stalk and used cryo-EM to show that this reversed the direction the motor points when bound to microtubules. This engineered motor moved robustly in the opposite direction to wild-type dynein showing that the angle of the stalk is a major determinant of directionality (Can et al 2019).<\/p>\n\n\n\n2D classes showing an engineered reverse direction dynein points toward the microtubule plus end.<\/figcaption><\/figure>\n<\/div>\n<\/div>\n\n\n\nConnecting dynein to cargos<\/h2>\n\n\n\n Our first work on cargos focused on BICD2, an adaptor known to link dynein to Golgi vesicles. We made fully recombinant human dynein and discovered that both BICD2 and dynactin are required for it to move over long distances (Schlager et al 2014). This unanticipated finding established the principle that cargo adaptors can activate dynein and suggested a simple way in which dynein is controlled by the cargo it carries.<\/p>\n\n\n\n
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In early 2015 we published cryoEM structures of dynein\u2019s essential cofactor dynactin: a 23 subunit complex built around a filament of the actin relate protein Arp1. The work revealed the architecture of dynactin and explained how the length of the Arp1 filament is specified to be a defined length. We also addressed how dynactin binds dynein. Instead of forming a loosely coupled complex, as previously anticipated, we showed that dynein and dynactin form an intimate interaction with BICD2 acting as the glue that sticks the dynein tail to the dynactin filament (Urnavicius et al 2015). Subsequently we used EM to uncover how this interaction drives large rearrangements in dynein and how this activates its long distance movement (Zhang et al 2017).<\/p>\n<\/div>\n\n\n\n
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Model of dynactin based on our cryoEM structure<\/figcaption><\/figure>\n<\/div>\n<\/div>\n\n\n\n\n
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BICD2 is one of a growing family of cargo adaptors that can activate dynein movement. These adaptors contain a long (>200 amino acid) coiled coil but lack conserved sequence motifs. In 2018 we published cryoEM structures of dynein\/dynactin bound to two new adaptors: BICD-related-1 (BICDR1) and HOOK3 (Urnavicius et al 2018). These show both adaptors robustly recruit a second dynein to dynactin. The two dyneins lie side-by-side stabilizing them sufficiently to allow the determination of a high-resolution structure of the dynein tail. We also used single molecule motility assays to show that the two dyneins made the dynein\/dynactin complex significantly faster than when a single dynein is present.<\/p>\n<\/div>\n\n\n\n
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CryoEM map of dynein’s tail bound to dynactin and the adaptor BICD2.<\/figcaption><\/figure>\n<\/div>\n<\/div>\n\n\n\n<\/p>\n","protected":false},"excerpt":{"rendered":"
The dynein motor All dyneins contain a motor domain made of a ring of six AAA+ domains. The AAA+ family are a widespread class of ATPases that typically unfold or remodel other proteins.\u00a0 A major question was how dynein uses its AAA+ architecture to generate force toward the minus ends of microtubules.\u00a0 Work from Andrew […]<\/p>\n","protected":false},"author":26,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","footnotes":"","_links_to":"","_links_to_target":""},"class_list":{"0":"post-1021","1":"page","2":"type-page","3":"status-publish","5":"entry"},"featured_image_src":null,"featured_image_src_square":null,"_links":{"self":[{"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/pages\/1021","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/users\/26"}],"replies":[{"embeddable":true,"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/comments?post=1021"}],"version-history":[{"count":12,"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/pages\/1021\/revisions"}],"predecessor-version":[{"id":1065,"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/pages\/1021\/revisions\/1065"}],"wp:attachment":[{"href":"https:\/\/www2.mrc-lmb.cam.ac.uk\/groups\/cartera\/wp-json\/wp\/v2\/media?parent=1021"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
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