An intramolecular interaction between the FHA domain and a coiled coil negatively regulates the kinesin motor KIF1A

Lee, Jae-Ran; Shin, Hye‐Won; Choi, June-Seok; Ko, Jaewon; Kim, Se‐Ho; Lee, Hyun Woo; Kim, Karam; Rho, Seong-Hwan; Lee, Jun Hyuck; Song, Hye-Eun; Eom, Soo Hyun; Kim, Eunjoon

doi:10.1038/sj.emboj.7600164

Cited by 68 publications

(83 citation statements)

References 37 publications

(56 reference statements)

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“…OSM-3, a kinesin-2 protein of C. elegans, is also an inactive motor in its native form in vitro, but can be turned on to an active motor by attaching an artificial cargo at the tail domain or introducing a point mutation in the stalk region that disrupts its intramolecular fold (9). KIF1A, a kinesin-3 protein, also participates in intramolecular interactions and mutations designed to disrupt these interactions convert it to an active motor in neuronal cells (11). Together, our results provide a proof of principle evidence that may govern generally conserved regulatory mechanisms for the activation of kinesins across species.…”

Section: Binding Of a Specific Segment Of Hdlg Activates Gakinmentioning

confidence: 99%

The Effector Domain of Human Dlg Tumor Suppressor Acts as a Switch That Relieves Autoinhibition of Kinesin-3 Motor GAKIN/KIF13B

2007

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Activity of motor proteins must be tightly regulated in the cells, in order to prevent unnecessary energy consumption and to maintain proper distribution of cellular components. Loading of the cargo molecule is one likely mechanism to activate an inactive motor. Here, we report that the activity of the kinesin-3 motor protein, GAKIN, is regulated by the direct binding of its protein cargo, human Discs large (hDlg) tumor suppressor. Recombinant GAKIN exhibits potent microtubule gliding activity but has little microtubule-stimulated ATPase activity in solution, suggesting that it exists in an auto-inhibitory form. In vitro binding measurements revealed that defined segments of GAKIN, particularly the MAGUK binding stalk (MBS) domain and the motor domain, mediate intramolecular interactions to confer globular protein conformation. Direct binding of the SH3-I3-GUK module of hDlg to the MBS domain of GAKIN activates the microtubule-stimulated ATPase activity of GAKIN by ~10 fold. We propose that the cargo-mediated regulation of motor activity constitutes a general paradigm for the activation of kinesins.Kinesin motors mediate the intracellular transport of cargo molecules along microtubule tracks (1). The mechanism that regulates the activation of kinesin motors remains an issue of fundamental importance (2,3). It has been reported that the full length conventional kinesin, kinesin-1, has very little microtubule-stimulated ATPase activity in solution as compared to its shorter motor domain constructs (4,5). The intramolecular interactions mediated by the motor and tail domains keep the kinesin-1 in a compactly-folded inhibitory state (6)(7)(8). It is well established that the mechanical attachment of the inactive full length motor to a glass surface or latex beads is sufficient to transform the inactive motor to an active state, presumably by disrupting its inhibitory conformation (8,9). Therefore, it is reasonable to speculate that this mode of self-inhibition of kinesin-1 is the general feature conserved in all kinesin-like proteins to keep their motor activity regulated in vivo.GAKIN/KIF13B (10) belongs to the kinesin-3 family. The family also includes KIF1A/ Unc104, which is responsible for the transport of synaptic vesicles (11). Our previous studies have identified two cargo molecules for GAKIN; the membrane-associated guanylate kinase homologue (MAGUK) scaffolding protein hDlg/SAP97 (12) and a PIP 3 binding protein termed PIP 3 BP (13). Both MAGUKs and PIP 3 have been implicated in the regulation of cell polarity pathways, and their subcellular localization is considered to be important for their functions in vivo. Recently, we have shown that the transport of PIP 3 containing vesicles to the distal * To whom correspondence should be addressed: A.H.C. (e-mail: chishti@uic.edu, phone: 312-355-1293A.H.C. (e-mail: chishti@uic.edu, phone: 312-355- , Fax: 312-355-1297 Recombinant protein expression in bacteriaGST fusion proteins were expressed in DH5α and purified using glutathione-Sepharose 4B (Pharmacia...

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Section: Binding Of a Specific Segment Of Hdlg Activates Gakinmentioning

confidence: 99%

The Effector Domain of Human Dlg Tumor Suppressor Acts as a Switch That Relieves Autoinhibition of Kinesin-3 Motor GAKIN/KIF13B

2007

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“…Cargo-binding proteins and proteins mediating the binding of cargoes to the tail region of kinesins are presumed to activate the kinesin motor (11,12). Pausing of the motor preventing unwanted activity depends on regulatory proteins or internal interactions within the kinesin molecule (13)(14)(15)(16)(17)(18). The regulatory proteins modulating activity of the kinesins in response to different cellular signals are still being identified.…”

mentioning

confidence: 99%

Structure of the complex of a mitotic kinesin with its calcium binding regulator

Vinogradova

Malanina

Reddy

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Much of the transport, tension, and movement in mitosis depends on kinesins, the ATP-powered microtubule-based motors. We report the crystal structure of a kinesin complex, the mitotic kinesin KCBP bound to its principal regulator KIC. Shown to be a Ca 2؉ sensor, KIC works as an allosteric trap. Extensive intermolecular interactions with KIC stabilize kinesin in its ADP-bound conformation. A critical component of the kinesin motile mechanism, called the neck mimic, switches its association from kinesin to KIC, stalling the motor. KIC denies access of the motor to its track by steric interference. Two major features of this regulation, allosteric trapping and steric blocking, are likely to be general for all kinesins.EF-hand ͉ motor ͉ calmodulin ͉ regulation

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“…Kinesin-3 motor proteins contain two characteristic FHA (forkhead-associated) and PH (pleckstrin homology) domains, in addition to the strongly conserved N-terminal motor domain. FHA domains are involved in phosphorylation-dependent protein-protein interactions and signaling (3,4), while the PH domain was shown to enable specific binding to phosphatidylinositol-4,5-bisphosphate in membranes (5).…”

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confidence: 99%

Neurospora crassa NKIN2, a Kinesin-3 Motor, Transports Early Endosomes and Is Required for Polarized Growth

et al. 2013

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bBiological motors are molecular nanomachines, which convert chemical energy into mechanical forces. The combination of mechanoenzymes with structural components, such as the cytoskeleton, enables eukaryotic cells to overcome entropy, generate molecular gradients, and establish polarity. Hyphae of filamentous fungi are among the most polarized cells, and polarity defects are most obvious. Here, we studied the role of the kinesin-3 motor, NKIN2, in Neurospora crassa. We found that NKIN2 localizes as fast-moving spots in the cytoplasm of mature hyphae. To test whether the spots represented early endosomes, the Rab5 GTPase YPT52 was used as an endosomal marker. NKIN2 colocalized with YPT52. Deletion of nkin2 caused strongly reduced endosomal movement. Combined, these results confirm the involvement of NKIN2 in early endosome transport. Introduction of a rigor mutation into NKIN2 labeled with green fluorescent protein (GFP) resulted in decoration of microtubules. Interestingly, NKIN2rigor was associated with a subpopulation of microtubules, as had been shown earlier for the Aspergillus nidulans orthologue UncA. Other kinesins did not show this specificity.

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An intramolecular interaction between the FHA domain and a coiled coil negatively regulates the kinesin motor KIF1A

Cited by 68 publications

References 37 publications

The Effector Domain of Human Dlg Tumor Suppressor Acts as a Switch That Relieves Autoinhibition of Kinesin-3 Motor GAKIN/KIF13B

The Effector Domain of Human Dlg Tumor Suppressor Acts as a Switch That Relieves Autoinhibition of Kinesin-3 Motor GAKIN/KIF13B

Structure of the complex of a mitotic kinesin with its calcium binding regulator

Neurospora crassa NKIN2, a Kinesin-3 Motor, Transports Early Endosomes and Is Required for Polarized Growth

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