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1.
Cell ; 186(12): 2531-2543.e11, 2023 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-37295401

RESUMO

RNA editing is a widespread epigenetic process that can alter the amino acid sequence of proteins, termed "recoding." In cephalopods, most transcripts are recoded, and recoding is hypothesized to be an adaptive strategy to generate phenotypic plasticity. However, how animals use RNA recoding dynamically is largely unexplored. We investigated the function of cephalopod RNA recoding in the microtubule motor proteins kinesin and dynein. We found that squid rapidly employ RNA recoding in response to changes in ocean temperature, and kinesin variants generated in cold seawater displayed enhanced motile properties in single-molecule experiments conducted in the cold. We also identified tissue-specific recoded squid kinesin variants that displayed distinct motile properties. Finally, we showed that cephalopod recoding sites can guide the discovery of functional substitutions in non-cephalopod kinesin and dynein. Thus, RNA recoding is a dynamic mechanism that generates phenotypic plasticity in cephalopods and can inform the characterization of conserved non-cephalopod proteins.


Assuntos
Cefalópodes , Dineínas , Animais , Dineínas/genética , Dineínas/metabolismo , Cinesinas/genética , Cinesinas/metabolismo , RNA/metabolismo , Cefalópodes/genética , Cefalópodes/metabolismo , Proteínas/metabolismo , Microtúbulos/metabolismo , Proteínas dos Microtúbulos , Miosinas/metabolismo
2.
Cell ; 186(12): 2544-2555.e13, 2023 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-37295402

RESUMO

In poikilotherms, temperature changes challenge the integration of physiological function. Within the complex nervous systems of the behaviorally sophisticated coleoid cephalopods, these problems are substantial. RNA editing by adenosine deamination is a well-positioned mechanism for environmental acclimation. We report that the neural proteome of Octopus bimaculoides undergoes massive reconfigurations via RNA editing following a temperature challenge. Over 13,000 codons are affected, and many alter proteins that are vital for neural processes. For two highly temperature-sensitive examples, recoding tunes protein function. For synaptotagmin, a key component of Ca2+-dependent neurotransmitter release, crystal structures and supporting experiments show that editing alters Ca2+ binding. For kinesin-1, a motor protein driving axonal transport, editing regulates transport velocity down microtubules. Seasonal sampling of wild-caught specimens indicates that temperature-dependent editing occurs in the field as well. These data show that A-to-I editing tunes neurophysiological function in response to temperature in octopus and most likely other coleoids.


Assuntos
Octopodiformes , Proteoma , Animais , Proteoma/metabolismo , Octopodiformes/genética , Edição de RNA , Temperatura , Sistema Nervoso/metabolismo , Adenosina Desaminase/metabolismo , RNA/metabolismo
3.
Annu Rev Cell Dev Biol ; 38: 155-178, 2022 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-35905769

RESUMO

Eukaryotic cells across the tree of life organize their subcellular components via intracellular transport mechanisms. In canonical transport, myosin, kinesin, and dynein motor proteins interact with cargos via adaptor proteins and move along filamentous actin or microtubule tracks. In contrast to this canonical mode, hitchhiking is a newly discovered mode of intracellular transport in which a cargo attaches itself to an already-motile cargo rather than directly associating with a motor protein itself. Many cargos including messenger RNAs, protein complexes, and organelles hitchhike on membrane-bound cargos. Hitchhiking-like behaviors have been shown to impact cellular processes including local protein translation, long-distance signaling, and organelle network reorganization. Here, we review instances of cargo hitchhiking in fungal, animal, and plant cells and discuss the potential cellular and evolutionary importance of hitchhiking in these different contexts.


Assuntos
Dineínas , Cinesinas , Actinas/metabolismo , Animais , Dineínas/genética , Dineínas/metabolismo , Cinesinas/genética , Microtúbulos/genética , Microtúbulos/metabolismo , Miosinas/genética , Miosinas/metabolismo , Células Vegetais/metabolismo
4.
Cell ; 173(4): 839-850.e18, 2018 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-29628142

RESUMO

Maize abnormal chromosome 10 (Ab10) encodes a classic example of true meiotic drive that converts heterochromatic regions called knobs into motile neocentromeres that are preferentially transmitted to egg cells. Here, we identify a cluster of eight genes on Ab10, called the Kinesin driver (Kindr) complex, that are required for both neocentromere motility and preferential transmission. Two meiotic drive mutants that lack neocentromere activity proved to be kindr epimutants with increased DNA methylation across the entire gene cluster. RNAi of Kindr induced a third epimutant and corresponding loss of meiotic drive. Kinesin gliding assays and immunolocalization revealed that KINDR is a functional minus-end-directed kinesin that localizes specifically to knobs containing 180 bp repeats. Sequence comparisons suggest that Kindr diverged from a Kinesin-14A ancestor ∼12 mya and has driven the accumulation of > 500 Mb of knob repeats and affected the segregation of thousands of genes linked to knobs on all 10 chromosomes.


Assuntos
Centrômero/metabolismo , Cinesinas/metabolismo , Meiose , Proteínas de Plantas/metabolismo , Zea mays/metabolismo , Centrômero/genética , Cromossomos de Plantas , Evolução Molecular , Haplótipos , Hibridização in Situ Fluorescente , Cinesinas/antagonistas & inibidores , Cinesinas/classificação , Cinesinas/genética , Modelos Genéticos , Mutagênese , Filogenia , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/classificação , Proteínas de Plantas/genética , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Sequenciamento Completo do Genoma , Zea mays/genética
5.
Cell ; 175(3): 796-808.e14, 2018 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-30340043

RESUMO

During cell division, mitotic motors organize microtubules in the bipolar spindle into either polar arrays at the spindle poles or a "nematic" network of aligned microtubules at the spindle center. The reasons for the distinct self-organizing capacities of dynamic microtubules and different motors are not understood. Using in vitro reconstitution experiments and computer simulations, we show that the human mitotic motors kinesin-5 KIF11 and kinesin-14 HSET, despite opposite directionalities, can both organize dynamic microtubules into either polar or nematic networks. We show that in addition to the motor properties the natural asymmetry between microtubule plus- and minus-end growth critically contributes to the organizational potential of the motors. We identify two control parameters that capture system composition and kinetic properties and predict the outcome of microtubule network organization. These results elucidate a fundamental design principle of spindle bipolarity and establish general rules for active filament network organization.


Assuntos
Cinesinas/metabolismo , Microtúbulos/metabolismo , Simulação de Dinâmica Molecular , Fuso Acromático/metabolismo , Animais , Humanos , Cinesinas/química , Microtúbulos/química , Células Sf9 , Fuso Acromático/química , Spodoptera
6.
Cell ; 167(2): 539-552.e14, 2016 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-27716509

RESUMO

Microtubule-organizing centers (MTOCs) nucleate microtubules that can grow autonomously in any direction. To generate bundles of parallel microtubules originating from a single MTOC, the growth of multiple microtubules needs to coordinated, but the underlying mechanism is unknown. Here, we show that a conserved two-component system consisting of the plus-end tracker EB1 and the minus-end-directed molecular motor Kinesin-14 is sufficient to promote parallel microtubule growth. The underlying mechanism relies on the ability of Kinesin-14 to guide growing plus ends along existing microtubules. The generality of this finding is supported by yeast, Drosophila, and human EB1/Kinesin-14 pairs. We demonstrate that plus-end guiding involves a directional switch of the motor due to a force applied via a growing microtubule end. The described mechanism can account for the generation of parallel microtubule networks required for a broad range of cellular functions such as spindle assembly or cell polarization.


Assuntos
Proteínas de Ciclo Celular/metabolismo , DNA Helicases/metabolismo , Cinesinas/metabolismo , Proteínas dos Microtúbulos/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Proteínas Motores Moleculares/metabolismo , Proteínas Oncogênicas/metabolismo , RNA Helicases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Animais , Drosophila melanogaster , Humanos , Fenômenos Mecânicos
7.
Genes Dev ; 37(5-6): 191-203, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36859340

RESUMO

Subcellular localization of messenger RNA (mRNA) is a widespread phenomenon that can impact the regulation and function of the encoded protein. In nonneuronal cells, specific mRNAs localize to cell protrusions, and proper mRNA localization is required for cell migration. However, the mechanisms by which mRNA localization regulates protein function in this setting remain unclear. Here, we examined the functional consequences of localization of the mRNA encoding KIF1C. KIF1C is a kinesin motor protein required for cell migration and mRNA trafficking, including trafficking of its own mRNA. We show that Kif1c mRNA localization does not regulate KIF1C's protein abundance, distribution, or ability to traffic other mRNAs. Conversely, Kif1c mRNA localization to protrusions is required for directed cell migration. We used mass spectrometry to identify binding partners of endogenous KIF1C, which revealed dramatic dysregulation of the number and identity of KIF1C interactors in response to Kif1c mRNA mislocalization. These results therefore uncovered a mechanistic connection between mRNA localization to cell protrusions and the specificity of protein-protein interactions. We anticipate that this mechanism is not limited to Kif1c and is likely to be a general principle that impacts the functions of proteins encoded by protrusion-enriched mRNAs in nonneuronal cells.


Assuntos
Cinesinas , Proteínas , RNA Mensageiro/metabolismo , Proteínas/metabolismo , Cinesinas/genética , Cinesinas/metabolismo , Dineínas/metabolismo , Movimento Celular/genética
8.
Genes Dev ; 37(5-6): 137-139, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36889919

RESUMO

Distinct subcellular localizations of mRNAs have been described across a wide variety of cell types. While common themes emerge for neuronal cells, functional roles of mRNA localization in space and time are much less understood in nonneuronal cells. Emerging areas of interest are cell models with protrusions, often linked with cell mobility in cancer systems. In this issue of Genes & Development, Norris and Mendell (pp. 191-203) systematically investigate a link between mRNA localization to cell protrusions in a mouse melanoma cell system and a mechanistic link to downstream consequences for cell mobility. The study first identifies a model mRNA of interest in an unbiased way that exhibits a set of phenotypes associated with cell mobility. The candidate mRNA that fulfills all requirements is Kif1c mRNA. Further systematic investigation links Kif1c mRNA localization to assembly of a protein-protein network on the KIF1C protein itself. What's clear is that this work will inspire a further mechanistic dissection of the Kif1c mRNA/KIF1C protein interplay in this important nonneuronal model cell system. More broadly, this work suggests that a broad set of model mRNAs should be investigated to understand mRNA dynamics and downstream functional consequences across a variety of cell models.


Assuntos
Cinesinas , Proteínas , Camundongos , Animais , Cinesinas/genética , Cinesinas/metabolismo , Ligação Proteica , RNA Mensageiro/metabolismo , Proteínas/metabolismo , Movimento Celular/genética
9.
Annu Rev Cell Dev Biol ; 32: 173-195, 2016 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-27362645

RESUMO

Objects are commonly moved within the cell by either passive diffusion or active directed transport. A third possibility is advection, in which objects within the cytoplasm are moved with the flow of the cytoplasm. Bulk movement of the cytoplasm, or streaming, as required for advection, is more common in large cells than in small cells. For example, streaming is observed in elongated plant cells and the oocytes of several species. In the Drosophila oocyte, two stages of streaming are observed: relatively slow streaming during mid-oogenesis and streaming that is approximately ten times faster during late oogenesis. These flows are implicated in two processes: polarity establishment and mixing. In this review, I discuss the underlying mechanism of streaming, how slow and fast streaming are differentiated, and what we know about the physiological roles of the two types of streaming.


Assuntos
Corrente Citoplasmática , Drosophila/citologia , Oócitos/citologia , Animais , Polaridade Celular , Oogênese
10.
Genes Dev ; 35(13-14): 976-991, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34140355

RESUMO

Kinesin-1 carries cargos including proteins, RNAs, vesicles, and pathogens over long distances within cells. The mechanochemical cycle of kinesins is well described, but how they establish cargo specificity is not fully understood. Transport of oskar mRNA to the posterior pole of the Drosophila oocyte is mediated by Drosophila kinesin-1, also called kinesin heavy chain (Khc), and a putative cargo adaptor, the atypical tropomyosin, aTm1. How the proteins cooperate in mRNA transport is unknown. Here, we present the high-resolution crystal structure of a Khc-aTm1 complex. The proteins form a tripartite coiled coil comprising two in-register Khc chains and one aTm1 chain, in antiparallel orientation. We show that aTm1 binds to an evolutionarily conserved cargo binding site on Khc, and mutational analysis confirms the importance of this interaction for mRNA transport in vivo. Furthermore, we demonstrate that Khc binds RNA directly and that it does so via its alternative cargo binding domain, which forms a positively charged joint surface with aTm1, as well as through its adjacent auxiliary microtubule binding domain. Finally, we show that aTm1 plays a stabilizing role in the interaction of Khc with RNA, which distinguishes aTm1 from classical motor adaptors.


Assuntos
Proteínas de Drosophila , Cinesinas , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Cinesinas/genética , Microtúbulos/metabolismo , Transporte de RNA , RNA Mensageiro/metabolismo , Tropomiosina/metabolismo
11.
Genes Dev ; 35(13-14): 937-939, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-34210804

RESUMO

The prototypic and ubiquitous microtubule motor, kinesin-1, uses a variety of adaptor proteins to facilitate the selective transport of diverse cargo within the cell. These cargo adaptors bind to the motor complex through interactions with the kinesin light or heavy chains (KLCs or KHCs). In this issue of Genes & Development, Dimitrova-Paternoga et al. (pp. 976-991) present the first structural characterization of a KHC-cargo adaptor interface. They describe an antiparallel heterotrimeric coiled-coil complex between the carboxy tail of KHC and Tm1-I/C (aTm1), the atypical tropomyosin that is important for oskar mRNA transport in Drosophila oocytes. This interaction enhances direct binding between KHC and RNA. Their findings demonstrate the structural plasticity of the KHC tail as a platform for protein-protein interactions and reveal how a cargo adaptor protein can modify a motor-RNA interface to promote transport.


Assuntos
Proteínas de Drosophila , Cinesinas , Animais , Drosophila/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Cinesinas/genética , Microtúbulos/metabolismo , RNA/metabolismo
12.
EMBO J ; 43(13): 2606-2635, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38806659

RESUMO

Microtubule-based kinesin motor proteins are crucial for intracellular transport, but their hyperactivation can be detrimental for cellular functions. This study investigated the impact of a constitutively active ciliary kinesin mutant, OSM-3CA, on sensory cilia in C. elegans. Surprisingly, we found that OSM-3CA was absent from cilia but underwent disposal through membrane abscission at the tips of aberrant neurites. Neighboring glial cells engulf and eliminate the released OSM-3CA, a process that depends on the engulfment receptor CED-1. Through genetic suppressor screens, we identified intragenic mutations in the OSM-3CA motor domain and mutations inhibiting the ciliary kinase DYF-5, both of which restored normal cilia in OSM-3CA-expressing animals. We showed that conformational changes in OSM-3CA prevent its entry into cilia, and OSM-3CA disposal requires its hyperactivity. Finally, we provide evidence that neurons also dispose of hyperactive kinesin-1 resulting from a clinic variant associated with amyotrophic lateral sclerosis, suggesting a widespread mechanism for regulating hyperactive kinesins.


Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Cílios , Cinesinas , Neuroglia , Animais , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/genética , Cinesinas/metabolismo , Cinesinas/genética , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Neuroglia/metabolismo , Cílios/metabolismo , Neurônios/metabolismo , Mutação , Esclerose Lateral Amiotrófica/metabolismo , Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/patologia
13.
EMBO J ; 43(7): 1244-1256, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38424239

RESUMO

During mitosis, motor proteins and microtubule-associated protein organize the spindle apparatus by cross-linking and sliding microtubules. Kinesin-5 plays a vital role in spindle formation and maintenance, potentially inducing twist in the spindle fibers. The off-axis power stroke of kinesin-5 could generate this twist, but its implications in microtubule organization remain unclear. Here, we investigate 3D microtubule-microtubule sliding mediated by the human kinesin-5, KIF11, and found that the motor caused right-handed helical motion of anti-parallel microtubules around each other. The sidestepping ratio increased with reduced ATP concentration, indicating that forward and sideways stepping of the motor are not strictly coupled. Further, the microtubule-microtubule distance (motor extension) during sliding decreased with increasing sliding velocity. Intriguingly, parallel microtubules cross-linked by KIF11 orbited without forward motion, with nearly full motor extension. Altering the length of the neck linker increased the forward velocity and pitch of microtubules in anti-parallel overlaps. Taken together, we suggest that helical motion and orbiting of microtubules, driven by KIF11, contributes to flexible and context-dependent filament organization, as well as torque regulation within the mitotic spindle.


Assuntos
Cinesinas , Microtúbulos , Humanos , Cinesinas/metabolismo , Microtúbulos/metabolismo , Fuso Acromático/fisiologia , Proteínas Associadas aos Microtúbulos/metabolismo , Mitose
14.
EMBO J ; 43(15): 3192-3213, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38898313

RESUMO

In cells, mRNAs are transported to and positioned at subcellular areas to locally regulate protein production. Recent studies have identified the kinesin-3 family member motor protein KIF1C as an RNA transporter. However, it is not clear how KIF1C interacts with RNA molecules. Here, we show that the KIF1C C-terminal tail domain contains an intrinsically disordered region (IDR) that drives liquid-liquid phase separation (LLPS). KIF1C forms dynamic puncta in cells that display physical properties of liquid condensates and incorporate RNA molecules in a sequence-selective manner. Endogenous KIF1C forms condensates in cellular protrusions, where mRNAs are enriched in an IDR-dependent manner. Purified KIF1C tail constructs undergo LLPS in vitro at near-endogenous nM concentrations and in the absence of crowding agents and can directly recruit RNA molecules. Overall, our work uncovers an intrinsic correlation between the LLPS activity of KIF1C and its role in mRNA positioning. In addition, the LLPS activity of KIF1C's tail represents a new mode of motor-cargo interaction that extends our current understanding of cytoskeletal motor proteins.


Assuntos
Cinesinas , RNA Mensageiro , Humanos , Proteínas Intrinsicamente Desordenadas/metabolismo , Proteínas Intrinsicamente Desordenadas/genética , Proteínas Intrinsicamente Desordenadas/química , Cinesinas/metabolismo , Cinesinas/genética , Separação de Fases , RNA Mensageiro/metabolismo , RNA Mensageiro/genética
15.
Genes Dev ; 34(17-18): 1110-1112, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32873577

RESUMO

Maize heterochromatic knobs cheat female meiosis by forming neocentromeres that bias their segregation into the future egg cell. In this issue of Genes & Development, Swentowsky and colleagues (pp. 1239-1251) show that two types of knobs, those composed of 180-bp and TR1 sequences, recruit their own novel and divergent kinesin-14 family members to form neocentromeres.


Assuntos
Genoma de Planta , Zea mays/genética , Centrômero/genética , Genoma de Planta/genética , Cinesinas/genética , Cinesinas/metabolismo , Meiose/genética
16.
Genes Dev ; 34(17-18): 1239-1251, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32820038

RESUMO

A maize chromosome variant called abnormal chromosome 10 (Ab10) converts knobs on chromosome arms into neocentromeres, causing their preferential segregation to egg cells in a process known as meiotic drive. We previously demonstrated that the gene Kinesin driver (Kindr) on Ab10 encodes a kinesin-14 required to mobilize neocentromeres made up of the major tandem repeat knob180. Here we describe a second kinesin-14 gene, TR-1 kinesin (Trkin), that is required to mobilize neocentromeres made up of the minor tandem repeat TR-1. Trkin lies in a 4-Mb region of Ab10 that is not syntenic with any other region of the maize genome and shows extraordinary sequence divergence from Kindr and other kinesins in plants. Despite its unusual structure, Trkin encodes a functional minus end-directed kinesin that specifically colocalizes with TR-1 in meiosis, forming long drawn out neocentromeres. TRKIN contains a nuclear localization signal and localizes to knobs earlier in prophase than KINDR. The fact that TR-1 repeats often co-occur with knob180 repeats suggests that the current role of the TRKIN/TR-1 system is to facilitate the meiotic drive of the KINDR/knob180 system.


Assuntos
Centrômero/genética , Centrômero/metabolismo , Cinesinas/genética , Cinesinas/metabolismo , Zea mays/genética , Zea mays/metabolismo , Cromossomos de Plantas/genética , Genes de Plantas/genética , Meiose , Modelos Genéticos , Transporte Proteico/genética
17.
EMBO J ; 42(10): e111559, 2023 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-37038978

RESUMO

Various cancer types exhibit characteristic and recurrent aneuploidy patterns. The origins of these cancer type-specific karyotypes are still unknown, partly because introducing or eliminating specific chromosomes in human cells still poses a challenge. Here, we describe a novel strategy to induce mis-segregation of specific chromosomes in different human cell types. We employed Tet repressor or nuclease-dead Cas9 to link a microtubule minus-end-directed kinesin (Kinesin14VIb) from Physcomitrella patens to integrated Tet operon repeats and chromosome-specific endogenous repeats, respectively. By live- and fixed-cell imaging, we observed poleward movement of the targeted loci during (pro)metaphase. Kinesin14VIb-mediated pulling forces on the targeted chromosome were counteracted by forces from kinetochore-attached microtubules. This tug-of-war resulted in chromosome-specific segregation errors during anaphase and revealed that spindle forces can heavily stretch chromosomal arms. By single-cell whole-genome sequencing, we established that kinesin-induced targeted mis-segregations predominantly result in chromosomal arm aneuploidies after a single cell division. Our kinesin-based strategy opens the possibility to investigate the immediate cellular responses to specific aneuploidies in different cell types; an important step toward understanding how tissue-specific aneuploidy patterns evolve.


Assuntos
Cinesinas , Fuso Acromático , Humanos , Cinesinas/genética , Cinesinas/metabolismo , Fuso Acromático/genética , Fuso Acromático/metabolismo , Cinetocoros/metabolismo , Microtúbulos/metabolismo , Segregação de Cromossomos , Anáfase , Aneuploidia
18.
EMBO J ; 42(21): e113647, 2023 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-37592895

RESUMO

During mitosis, spindle architecture alters as chromosomes segregate into daughter cells. The microtubule crosslinker protein regulator of cytokinesis 1 (PRC1) is essential for spindle stability, chromosome segregation and completion of cytokinesis, but how it recruits motors to the central spindle to coordinate the segregation of chromosomes is unknown. Here, we combine structural and cell biology approaches to show that the human CENP-E motor, which is essential for chromosome capture and alignment by microtubules, binds to PRC1 through a conserved hydrophobic motif. This binding mechanism is also used by Kinesin-4 Kif4A:PRC1. Using in vitro reconstitution, we demonstrate that CENP-E slides antiparallel PRC1-crosslinked microtubules. We find that the regulation of CENP-E -PRC1 interaction is spatially and temporally coupled with relocalization to overlapping microtubules in anaphase. Finally, we demonstrate that the PRC1-microtubule motor interaction is essential in anaphase to control chromosome partitioning, retain central spindle integrity and ensure cytokinesis. Taken together our findings reveal the molecular basis for the cell cycle regulation of motor-PRC1 complexes to couple chromosome segregation and cytokinesis.


Assuntos
Citocinese , Cinesinas , Humanos , Citocinese/fisiologia , Cinesinas/genética , Cinesinas/metabolismo , Fosforilação , Fuso Acromático/metabolismo , Mitose , Proteínas de Ciclo Celular/metabolismo , Microtúbulos/metabolismo
19.
EMBO J ; 42(16): e112812, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37403793

RESUMO

Intracellular organelle organization is conserved in eukaryotic cells and is primarily achieved through active transport by motor proteins along the microtubule cytoskeleton. Microtubule post-translational modifications (PTMs) can contribute to microtubule diversity and differentially regulate motor-mediated transport. Here, we show that centrosome amplification, commonly observed in cancer and shown to promote aneuploidy and invasion, induces a global change in organelle positioning towards the cell periphery and facilitates nuclear migration through confined spaces. This reorganization requires kinesin-1 and is analogous to the loss of dynein. Cells with amplified centrosomes display increased levels of acetylated tubulin, a PTM that could enhance kinesin-1-mediated transport. Depletion of α-tubulin acetyltransferase 1 (αTAT1) to block tubulin acetylation rescues the displacement of centrosomes, mitochondria, and vimentin but not Golgi or endosomes. Analyses of the distribution of total and acetylated microtubules indicate that the polarized distribution of modified microtubules, rather than levels alone, plays an important role in the positioning of specific organelles, such as the centrosome. We propose that increased tubulin acetylation differentially impacts kinesin-1-mediated organelle displacement to regulate intracellular organization.


Assuntos
Cinesinas , Tubulina (Proteína) , Tubulina (Proteína)/metabolismo , Cinesinas/genética , Cinesinas/metabolismo , Acetilação , Microtúbulos/metabolismo , Centrossomo/metabolismo , Dineínas/metabolismo , Processamento de Proteína Pós-Traducional
20.
EMBO J ; 42(5): e112101, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36636822

RESUMO

Tubulin posttranslational modifications have been predicted to control cytoskeletal functions by coordinating the molecular interactions between microtubules and their associating proteins. A prominent tubulin modification in neurons is polyglutamylation, the deregulation of which causes neurodegeneration. Yet, the underlying molecular mechanisms have remained elusive. Here, using in-vitro reconstitution, we determine how polyglutamylation generated by the two predominant neuronal polyglutamylases, TTLL1 and TTLL7, specifically modulates the activities of three major microtubule interactors: the microtubule-associated protein Tau, the microtubule-severing enzyme katanin and the molecular motor kinesin-1. We demonstrate that the unique modification patterns generated by TTLL1 and TTLL7 differentially impact those three effector proteins, thus allowing for their selective regulation. Given that our experiments were performed with brain tubulin from mouse models in which physiological levels and patterns of polyglutamylation were altered by the genetic knockout of the main modifying enzymes, our quantitative measurements provide direct mechanistic insight into how polyglutamylation could selectively control microtubule interactions in neurons.


Assuntos
Tubulina (Proteína) , Animais , Camundongos , Citoesqueleto/metabolismo , Cinesinas/metabolismo , Microtúbulos/metabolismo , Tubulina (Proteína)/metabolismo , Peptídeo Sintases , Proteínas Associadas aos Microtúbulos
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