Structural basis for isoform-specific kinesin-1 recognition of Y-acidic cargo adaptors

Pernigo, S.; Chegkazi, Magda S.; Yip, Yan Yan; Treacy, Conor; Glorani, Giulia; Hansen, Kjetil; Politis, Argyris; Bui, Soi; Dodding, Mark P.; Steiner, Roberto A.

doi:10.7554/elife.38362

Cited by 32 publications

(31 citation statements)

References 61 publications

(97 reference statements)

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“…The structure of the KBP–KIF15_MD6S complex shows how KBP binds the KIF15_MD6S via its concave face and undergoes subtle remodelling of its N-terminal domain to accommodate kinesin binding. This further reinforces the idea that the TPR-containing structures are not simply static scaffolds but can flexibly respond to ligand binding ( Pernigo et al, 2018 ). In contrast, the KIF15 motor domain undergoes a radical conformational change in forming a complex with KBP, in which helix α4, the major component of the motor’s tubulin-binding subdomain, is displaced from the main MD body by ~15 Å into the KBP concave face.…”

Section: Discussionsupporting

confidence: 79%

The mechanism of kinesin inhibition by kinesin-binding protein

Atherton

Hummel

Olieric

et al. 2020

eLife

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Subcellular compartmentalisation is necessary for eukaryotic cell function. Spatial and temporal regulation of kinesin activity is essential for building these local environments via control of intracellular cargo distribution. Kinesin binding protein (KBP) interacts with a subset of kinesins via their motor domains, inhibits their microtubule (MT) attachment and blocks their cellular function. However, its mechanisms of inhibition and selectivity have been unclear. Here we use cryo-electron microscopy to reveal the structure of KBP and of a KBP-kinesin motor domain complex. KBP is a TPR-containing, right-handed α-solenoid that sequesters the kinesin motor domain’s tubulin-binding surface, structurally distorting the motor domain and sterically blocking its MT attachment. KBP uses its α-solenoid concave face and edge loops to bind the kinesin motor domain, and selected structure-guided mutations disrupt KBP inhibition of kinesin transport in cells. The KBP-interacting motor domain surface contains motifs exclusively conserved in KBP-interacting kinesins, suggesting a basis for kinesin selectivity.

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Section: Discussionsupporting

confidence: 79%

The mechanism of kinesin inhibition by kinesin-binding protein

Atherton

Hummel

Olieric

et al. 2020

eLife

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“…5D-E). We additionally tested an orthogonal adapter protein, JIP1, which contains a Y-acidic motif, and has been shown to bind directly to the KLC TPR domain (Pernigo et al, 2018), and suggested to activate KHC motility in cell lysates (Fu and Holzbaur, 2013). We again did not observe any effect of JIP1 on the frequency of processive events with our purified kinesin tetramer (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A series of biochemical and structural studies have uncovered two consensus KLC binding sequences found within numerous cellular proteins that are thought to act as cargo-adapter molecules for kinesin (Cross and Dodding, 2019; Pernigo et al, 2013). The isolated C-terminal TPR domains of KLC bind to either W-acidic or Y-acidic motifs with relatively low affinity (~5 μM and 0.5 μM respectively, (Pernigo et al, 2018; Pernigo et al, 2013; Yip et al, 2016)), suggesting that the formation of kinesin-cargo complexes is also regulated by autoinhibition. Despite the low affinity in vitro , W-acidic peptides, or small molecule mimetic drugs (Randall et al, 2017), appear sufficient to drive kinesin activity within living cells, suggesting that cargo-adapter binding to the KLC could directly activate the autoinhibited motor.…”

Section: Introductionmentioning

confidence: 99%

Reconstitution of Kinesin-1 Activation

Chiba

Niwa

et al. 2021

Preprint

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Autoinhibition is an important regulatory mechanism for cytoskeletal motor proteins. Kinesin-1 (kinesin hereafter), the ubiquitous plus-end directed microtubule motor, is thought to be controlled by a complicated autoinihibition mechanism, but the molecular details remain unclear. Conformational changes mediated by intramolecular interactions between the C-terminal tail and N-terminal motor domains of the kinesin heavy chain (KHC) are proposed to be one facet of motor regulation. The dimeric KHC also binds two copies of the kinesin light chains (KLCs), which have been implicated in both autoinhibition and cargo-dependent activation of the tetrameric motor complex, although the precise mechanisms remain opaque. Using in vitro reconstitution, we show that the KLC strongly inhibits the kinesin-microtubule interaction via an independent mechanism from the tail-motor interaction within KHC. Kinesin cargo-adaptor proteins that bind KLC activated processive movement of the kinesin tetramer but the landing rate of these activated complexes remained low. The addition of MAP7, which specifically binds to the KHC, strongly enhanced activated motor motility by dramatically increasing the landing rate and processivity of the activated kinesin motors. Our results support a model whereby the activity of the kinesin tetramer is regulated by independent tail- and KLC-based inhibition mechanisms, and that cargo-adaptor binding to the KLC directly releases both of these inhibitions. However, we find that a third component, a non-motor MAP is required for robust activity of the activated motor. Thus, human kinesin activity is regulated by a two-factor mechanism comprised of intramolecular allosteric regulation, as well as intermolecular kinesin-adaptor and kinesin-MAP interactions.

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“…It should be noted that this pentapeptide partially overlaps a sequence (E 344 FYSE 348 ) that has the characteristics of a Y-acidic motif. The latter has recently been identified in the JIP1 protein and is, like the W-acidic motif, recognized by the KLC (Pernigo et al, 2018). However, the involvement of this sequence in the interaction between kinesin-1 and PipB2 is uncertain as the deletion of the pentapeptide, which partially covers this hypothesized Y-acidic motif, has only a partial effect on the interaction between PipB2 and KLC (Henry et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…In the cellular context, binding of the cargo protein JIP1 (also known as MAPK8IP1) to KLCs is not sufficient to activate kinesin-1 and requires additional interactions of KHCs with FEZ1 (Blasius et al, 2007). The TPR domain of KLC recognizes a peptide sequence composed of a tryptophan (Dodding et al, 2011) or a tyrosine (Pernigo et al, 2018) residue flanked by acid residues, sequences that have recently been named W-acidic and Y-acidic. The W-acidic motif originally characterized in vaccinia virus protein A36 is also found in many adaptor proteins such as SKIP, which contains two of them (Pernigo et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Regulation of kinesin-1 activity by the Salmonella enterica effectors PipB2 and SifA

Alberdi

Vergnes

Manneville

et al. 2020

Journal of Cell Science

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Salmonella enterica is an intracellular bacterial pathogen. The formation of its replication niche, which is composed of a vacuole associated with a network of membrane tubules, depends on the secretion of a set of bacterial effector proteins whose activities deeply modify the functions of the eukaryotic host cell. By recruiting and regulating the activity of the kinesin-1 molecular motor, Salmonella effectors PipB2 and SifA play an essential role in the formation of the bacterial compartments. In particular, they allow the formation of tubules from the vacuole and their extension along the microtubule cytoskeleton, and thus promote membrane exchanges and nutrient supply. We have developed in vitro and in cellulo assays to better understand the specific role played by these two effectors in the recruitment and regulation of kinesin-1. Our results reveal a specific interaction between the two effectors and indicate that, contrary to what studies on infected cells suggested, interaction with PipB2 is sufficient to relieve the autoinhibition of kinesin-1. Finally, they suggest the involvement of other Salmonella effectors in the control of the activity of this molecular motor.

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Structural basis for isoform-specific kinesin-1 recognition of Y-acidic cargo adaptors

Cited by 32 publications

References 61 publications

The mechanism of kinesin inhibition by kinesin-binding protein

The mechanism of kinesin inhibition by kinesin-binding protein

Reconstitution of Kinesin-1 Activation

Regulation of kinesin-1 activity by the Salmonella enterica effectors PipB2 and SifA

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