Mechanisms by Which Kinesin-5 Motors Perform Their Multiple Intracellular Functions

Pandey, Himanshu; Popov, Mary; Goldstein-Levitin, Alina; Gheber, Larisa

doi:10.3390/ijms22126420

Cited by 18 publications

(17 citation statements)

References 285 publications

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“…Kinesin-5 motor proteins (kinesin-5s) are essential for mitotic chromosome segregation, facilitating mitotic spindle assembly, maintenance, and elongation [reviewed in (1)(2)(3)(4)]. These proteins are bipolar kinesins, with two pairs of catalytic motor domains located on opposite sides of an elongated homotetrameric complex (5)(6)(7).…”

Section: Introductionmentioning

confidence: 99%

“…These proteins are bipolar kinesins, with two pairs of catalytic motor domains located on opposite sides of an elongated homotetrameric complex (5)(6)(7). By virtue of this bipolar structure, kinesin-5s cross-link the antiparallel microtubules (MTs) of the spindle, and by moving on the two crosslinked MTs, they provide the outwardly directed force essential for spindle pole separation during spindle assembly and anaphase B spindle elongation (1)(2)(3)(4).…”

Section: Introductionmentioning

confidence: 99%

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Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin

Singh,

Siegler,

Pandey

et al. 2024

Sci. Adv.

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Several kinesin-5 motors (kinesin-5s) exhibit bidirectional motility. The mechanism of such motility remains unknown. Bidirectional kinesin-5s share a long N-terminal nonmotor domain (NTnmd), absent in exclusively plus-end–directed kinesins. Here, we combined in vivo, in vitro, and cryo–electron microscopy (cryo-EM) studies to examine the impact of NTnmd mutations on the motor functions of the bidirectional kinesin-5, Cin8. We found that NTnmd deletion mutants exhibited cell viability and spindle localization defects. Using cryo-EM, we examined the structure of a microtubule (MT)-bound motor domain of Cin8, containing part of its NTnmd. Modeling and molecular dynamic simulations based on the cryo-EM map suggested that the NTnmd of Cin8 interacts with the C-terminal tail of β-tubulin. In vitro experiments on subtilisin-treated MTs confirmed this notion. Last, we showed that NTnmd mutants are defective in plus-end–directed motility in single-molecule and antiparallel MT sliding assays. These findings demonstrate that the NTnmd, common to bidirectional kinesin-5s, is critical for their bidirectional motility and intracellular functions.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin

Singh,

Siegler,

Pandey

et al. 2024

Sci. Adv.

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“…Our findings reveal profound up-regulation of genes encoding various kinesins in lung tissue of mice recovering from a sublethal PVM infection, as well as their diminished expression in PVMinfected mice treated with L. plantarum or mice treated with L. plantarum alone (Figure 5a). Among these are genes encoding kinesin-4 (encoded by Kif4) which promotes chromosome condensation together with kinesin-5 (encoded by Kif11) and kinesin-13 (encoded by Kif2), which establish and position and the mitotic spindle, respectively [65][66][67][68].…”

Section: Differential Expression Of Genes Encoding Kinesins (Kifs) Centromere Proteins (Cenps) and Aurora Kinasesmentioning

confidence: 99%

Differential Expression of Mitosis and Cell Cycle Regulatory Genes during Recovery from an Acute Respiratory Virus Infection

et al. 2021

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Acute respiratory virus infections can have profound and long-term effects on lung function that persist even after the acute responses have fully resolved. In this study, we examined gene expression by RNA sequencing in the lung tissue of wild-type BALB/c mice that were recovering from a sublethal infection with the pneumonia virus of mice (PVM), a natural rodent pathogen of the same virus family and genus as the human respiratory syncytial virus. We compared these responses to gene expression in PVM-infected mice treated with Lactobacillus plantarum, an immunobiotic agent that limits inflammation and averts the negative clinical sequelae typically observed in response to acute infection with this pathogen. Our findings revealed prominent differential expression of inflammation-associated genes as well as numerous genes and gene families implicated in mitosis and cell-cycle regulation, including cyclins, cyclin-dependent kinases, cell division cycle genes, E2F transcription factors, kinesins, centromere proteins, and aurora kinases, among others. Of particular note was the differential expression of the cell division cycle gene Cdc20b, which was previously identified as critical for the ex vivo differentiation of multi-ciliated cells. Collectively, these findings provided us with substantial insight into post-viral repair processes and broadened our understanding of the mechanisms underlying Lactobacillus-mediated protection.

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“…They can either stabilize or destabilize microtubule plus ends. One such interesting candidate is kinesin-5, also known as Eg5 or KIF11, which is essential for bipolar spindle assembly and regulation of spindle elongation (Saunders et al, 2007;Brust-Mascher et al, 2009;She et al, 2020;Pandey et al, 2021). It is a homotetrameric motor protein that can crosslink microtubules and slide them against each other by walking in both directions (Kapitein et al, 2005;Scholey et al, 2014).…”

Section: Microtubule Plus-end Regulationmentioning

confidence: 99%

Dynamic life of a microtubule

Rai¹

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Microtubules are one of the major types of cytoskeletal filaments in cells. They are very dynamic polymers composed of αβ-tubulin dimers arranged longitudinally in head-to-tail fashion as well as laterally to assemble 13-protofilament hollow cylindrical tubes. The incorporation of GTP-bound αβ-tubulin dimers generates a fast growing plus end exposing β-tubulin and a slow growing minus end exposing α-tubulin. In cells, microtubules are assembled de novo from a template, called γ-TuRC, which interacts with α-tubulin. Microtubules can either remain capped by γ-TuRC and anchored to the microtubule-organizing centers (MTOCs) or be released if they are cut by microtubule severing enzymes like katanin. The release of microtubules from MTOC generates free minus ends, which are then stabilized by minus-end binding proteins called CAMSAPs. However, the plus ends remain very dynamic and undergo transitions from growth to shrinkage, termed “catastrophes”, and the opposite transitions termed “rescues”. Numerous microtubule regulatory proteins act at the plus ends, minus ends and the microtubule shafts connecting the two ends to control the organization and density of cellular microtubule networks. In this thesis, we focused on each of these aspects and explored the dynamic life of microtubules by reconstituting these processes in vitro using purified proteins. We first focused on the birth and growth of microtubules. We reconstituted microtubule nucleation using purified γ-TuRC and microtubule regulatory proteins and showed that CDK5RAP2, CLASP2 and chTOG promoted microtubule nucleation from γ-TuRC. We discovered that CAMSAPs can bind to γ-TuRC-capped microtubule minus ends and displace γ-TuRC from these ends, generating free and stable microtubule minus ends. Furthermore, we found out that CDK5RAP2, but not CLASP2 or chTOG, can inhibit CAMSAP binding and microtubule release. We propose that the destiny of a microtubule depends on the type of protein complex that activates its nucleation. We then described a mechanism for stabilization of microtubule lattice by TRIM46, a neuronal protein, which can bundle parallel microtubules and promote microtubule rescues within these bundles. We also revealed that Ankyrin-G, a scaffold protein, can recruit TRIM46-stabilized microtubule bundles to the axonal membrane to drive the assembly of the axon initial segment in neurons. We also uncovered a new role of CLASP2 as a microtubule repair factor participating in microtubule maintenance. We demonstrated that CLASP2, an anti-catastrophe factor, can promote complete repair of damaged microtubule lattices by inhibiting microtubule depolymerization and promoting tube closure at the damage sites, causing lattice renewal. Finally, we described a three-protein module involving katanin, CAMSAPs, and WDR47 that can regulate microtubule polymer mass and minus-end stability. We showed that katanin can cut and amplify CAMSAP2/3-stabilized microtubule minus ends. WDR47 can inhibit the binding of katanin to CAMSAP2/3-stabilized minus ends and protect them from severing. The presence of WDR47 shifts the balance from microtubule amplification to minus-end growth regulation. To conclude, we obtained mechanistic insights into the regulation of microtubule nucleation, minus-end dynamics, lattice stabilization and maintenance, microtubule number and the interplay between microtubule regulatory proteins. These insights will help to understand how microtubule arrays are organized in cells.

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Mechanisms by Which Kinesin-5 Motors Perform Their Multiple Intracellular Functions

Cited by 18 publications

References 285 publications

Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin

Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin

Differential Expression of Mitosis and Cell Cycle Regulatory Genes during Recovery from an Acute Respiratory Virus Infection

Dynamic life of a microtubule

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