Comparison of transcriptional programs between undifferentiated (0 hr) and differentiated N2A-A5 (96 hr) cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE335602 | GEO | 2026/07/20
REPOSITORIES: GEO
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Project description:Kinesins are a superfamily of molecular motors that undertake ATP-dependent microtubule-based movement or regulate microtubule dynamics. Plasmodium species, which cause malaria and kill hundreds of thousands annually, encode ~9 kinesins in their genomes. Of these, two – kinesin-8B and kinesin-8X - are canonically classified as kinesin-8s, which in other eukaryotes typically modulate microtubule dynamics. Unexpectedly, Plasmodium kinesin-8B is required for development of the flagellated male gamete in the mosquito host, and its absence completely blocks parasite transmission. To understand the molecular basis of kinesin-8B’s essential role, we characterised the in vitro properties of the kinesin-8B motor domains from P. berghei and P. falciparum. Both motors drive plus-end directed ATP-dependent microtubule gliding, but also catalyse ATP-dependent microtubule depolymerisation. We determined the microtubule-bound structures of these motors using cryo-electron microscopy. P. berghei and P. falciparum kinesin-8B exhibit a very similar mode of microtubule interaction, in which Plasmodium-distinct sequences at the microtubule-kinesin interface influence motor function. Intriguingly, however, P. berghei kinesin-8B exhibits a non-canonical structural response to ATP analogue binding such that neck linker docking is not induced. Nevertheless, the neck linker region is absolutely required for motility and depolymerisation activities of these motors. Taken together, these data suggest that the mechanochemistry of Plasmodium kinesin-8Bs has been functionally tuned to efficiently contribute to flagella formation.
2022-12-05 | PXD037474 | Pride
Project description:Kinesins are motor proteins found in all eukaryotic lineages that move along microtubules to mediate Kinesins are motor proteins found in all eukaryotic lineages that move along microtubules to mediate cellular processes such as mitosis and intracellular transport. In trypanosomatids, the kinesin superfamily has undergone a prominent expansion, resulting in one of the most diverse kinesin repertoires that includes the two kinetoplastid-restricted families X1 and X2. Here, we characterize in Trypanosoma brucei TbKifX2A, an orphaned X2 kinesin. TbKifX2A tightly interacts with TbPH1, a kinesin-like protein with a likely inactive motor domain, a rarely reported occurrence. Both TbKifX2A and TbPH1 localize to the microtubule quartet (MtQ), a characteristic but poorly understood cytoskeletal structure that wraps around the flagellar pocket as it extends to the cell body anterior. The proximal proteome of TbPH1 revealed two other interacting proteins, the flagellar pocket protein FP45 and intriguingly another X2 kinesin, TbKifX2C. Simultaneous ablation of TbKifX2A/TbPH1 results in the depletion of FP45 and TbKifX2C and also an expansion of the flagellar pocket, among other morphological defects. TbKifX2A is the first motor protein to be localized to the MtQ. The observation that TbKifX2C also associates with the MtQ suggests that the X2 kinesin family may have co-evolved with the MtQ, both kinetoplastid-specific traits.cellular processes such as mitosis and intracellular transport. In trypanosomatids, the kinesin superfamily has undergone a prominent expansion, resulting in one of the most diverse kinesin repertoires that includes the two kinetoplastid-restricted families X1 and X2. Here, we characterize in Trypanosoma brucei TbKifX2A, an orphaned X2 kinesin. TbKifX2A tightly interacts with TbPH1, a kinesin-like protein with a likely inactive motor domain, a rarely reported occurrence. Both TbKifX2A and TbPH1 localize to the microtubule quartet (MtQ), a characteristic but poorly understood cytoskeletal structure that wraps around the flagellar pocket as it extends to the cell body anterior. The proximal proteome of TbPH1 revealed two other interacting proteins, the flagellar pocket protein FP45 and intriguingly another X2 kinesin, TbKifX2C. Simultaneous ablation of TbKifX2A/TbPH1 results in the depletion of FP45 and TbKifX2C and also an expansion of the flagellar pocket, among other morphological defects. TbKifX2A is the first motor protein to be localized to the MtQ. The observation that TbKifX2C also associates with the MtQ suggests that the X2 kinesin family may have co-evolved with the MtQ, both kinetoplastid-specific traits.
2022-06-30 | PXD033717 | Pride
Project description:Comparison of transcriptional programs between undifferentiated (0 hr) and differentiated N2A-A5 (96 hr) cells
| PRJNA1478971 | ENA
Project description:Purpose: Breast cancer is a genetically heterogenous disease with subtypes differing in prognosis and chemosensitivity. The basal-like breast cancer (BLBC) molecular subtype is associated with poorer outcomes, but is more responsive to taxane-based chemotherapy. We evaluated the role of kinesins, motor proteins interacting with microtubules, in influencing taxane resistance. Experimental Design: Kinesin (KIF) expression was studied in one local dataset comprising all taxane resistant breast cancers in relation to taxane resistance. Data in the NCI-60 cell line dataset (GSE5846) nd the MDACC dataset (GSE20194) is separately detailed. Results: In the local dataset, the kinesin KIF26B is overexpressed in taxane-resistant residual breast cancers post-chemotherapy. Conclusions: We show that kinesin overexpression correlates with taxane resistance in BLBC cell lines and tissue. Our results suggest a potential approach to overcoming taxane resistance through concurrent or sequential use of kinesin inhibitors, highlighting the ATP-binding domain as a drug development target. Kinesin (KIF) expression was studied in one local dataset comprising all taxane resistant breast cancers in relation to taxane resistance. Data in the NCI-60 cell line dataset (GSE5846) and the MDACC dataset (GSE20194) is separately detailed.
2012-12-01 | E-GEOD-22796 | biostudies-arrayexpress
Project description:Kinesins are motor proteins found in all eukaryotic lineages that move along microtubule tracks to mediate several cellular processes such as mitosis and intracellular transport of cargo. In trypanosomatids, the kinesin protein superfamily has undergone a prominent expansion, giving these protists one of the most diverse kinesin repertoires in eukaryotes. This has led to the emergence of two trypanosomatid-restricted groups of kinesins. Here, we characterize in Trypanosoma brucei TbKifX2, a hitherto orphaned kinesin that belongs to one of these groups. TbKifX2 tightly interacts with TbPH1, a kinesin-like protein named after a pleckstrin homology (PH) domain present within its carboxy terminal tail. TbKifX2 recruits TbPH1 to the microtubule quartet (MtQ), a characteristic cytoskeletal structure that runs adjacent to the flagellar attachment zone filament from the basal body to the anterior of the cell. The proximal proteome of TbPH1 has revealed four proteins that localize to structures found adjacent to the length of the MtQ, further confirming that the likely TbKifX1/TbPH1 heterodimer binds the MtQ along its whole length. Simultaneous ablation of both TbKifX2 and TbPH1 leads to the formation of prominent extrusions from the cell posterior. Thus, we have attributed a morphogenesis role to these two trypanosomatid-restricted proteins, and the unique cytoskeletal structure represented by the MtQ. We hypothesize that the putative TbKiX2/TbPH1 heterodimer may transport a cytokinesis auxiliary factor(s) along the MtQ to or from the T. brucei posterior. The cohort of proteins found in proximity to TbPH1 may represent one of these factors directly or be involved in their trafficking during cell division in trypanosomatids.
2022-06-30 | PXD025802 | Pride
Project description:RNA transport and local translation provide spatial control of gene expression, and RNA binding proteins (RBPs) act as critical adapters in this multi-step process. Muscleblind-like (MBNL) RNA binding proteins, implicated in Myotonic Dystrophy and cancer, localize RNAs to myoblast membranes and distal neurites, through unknown mechanisms. We found that MBNLs form motile and anchored granules in neurons and myoblasts, and selectively associate with kinesins Kif1bα and Kif1c through its zinc finger (ZnF) domains. Other RBPs with similar ZnFs also associate with these kinesins, implicating a motor-RBP specificity code. Live cell imaging and fractionation revealed that membrane anchoring is mediated through the unstructured carboxy-terminal tail of MBNL1. Both kinesin- and membrane-recruitment functions were reconstituted using MBNL-MS2 coat protein fusions; this approach, termed RBP Module Recruitment and Imaging (RBP-MRI), decouples RNA binding, kinesin recruitment, and membrane anchoring functions, while also establishing general strategies for studying multi-functional, modular domains of RBPs.
2022-07-09 | GSE207597 | GEO
Project description:Purpose: Breast cancer is a genetically heterogenous disease with subtypes differing in prognosis and chemosensitivity. The basal-like breast cancer (BLBC) molecular subtype is associated with poorer outcomes, but is more responsive to taxane-based chemotherapy. We evaluated the role of kinesins, motor proteins interacting with microtubules, in influencing taxane resistance. Experimental Design: Kinesin (KIF) expression was studied in one local dataset comprising all taxane resistant breast cancers in relation to taxane resistance. Data in the NCI-60 cell line dataset (GSE5846) nd the MDACC dataset (GSE20194) is separately detailed. Results: In the local dataset, the kinesin KIF26B is overexpressed in taxane-resistant residual breast cancers post-chemotherapy. Conclusions: We show that kinesin overexpression correlates with taxane resistance in BLBC cell lines and tissue. Our results suggest a potential approach to overcoming taxane resistance through concurrent or sequential use of kinesin inhibitors, highlighting the ATP-binding domain as a drug development target.
2012-12-01 | GSE22796 | GEO
Project description:Kinesin-1 is the first identified microtubule-based motor protein that drives anterograde intracellular transport of diverse cargoes in eukaryotic cells. Defects in kinesin-1 activity have been implicated in various human neurological disorders and intracellular pathogens such as bacteria and viruses hijack its activity. Despite its importance, the molecular mechanisms governing kinesin-1 activation remains poorly understood. Here, we report a 8.6-Å cryo-EM electron microscopy structure of the autoinhibited kinesin-1 heterotetramer. Our structure reveals a complex 36-nm head-to-tail assembly composed of two kinesin heavy chains (KHC) and two light chains (KLC), where the self-folded dimeric KHCs are stabilized by asymmetrically arranged KLCs tetratricopeptide repeat (TPR) domains. This state inhibits kinesin motility by constraining the dimeric motor domains in a configuration that is incompatible with motility. Notably, our structure also shows that the dimeric KLC TPR cargo-binding domains are occluded, providing a structural basis for the autoinhibition of both motor activity and cargo binding.In addition to functional studies, we also applied crosslinking mass spectrometry (XLMS) to validate the autoinhibitory model.
2026-06-03 | PXD073177 | Pride
Project description:The mitotic kinesin-like Protein 2 (MKLP2, KIF20A) is essential for the proper execution of cytokinesis and required to ‘passage’ the chromosomal passenger complex (CPC) from the chromosomes in (pro)metaphase to the spindle midzone and equatorial cortex in anaphase. Together with MKLP1 (KIF23) and MPP1 (KIF20B), MKLP2 forms the kinesin-6 subfamily of motor proteins. Whilst MKLP1 and MPP1 are both plus-end directed processive motors, the typical structure of the MKLP2 motor domain along with the extended neck-linker region, suggested that MKLP2 might not be able to function as a transport motor but is more likely to act as a microtubule bundler. This notion contradicts the prevailing hypothesis that MKLP2 can transport the CPC in anaphase, but appears to be in line with in vitro and in cellulo studies showing that recombinant MKLP2 efficiently bundles microtubules, and that knock-down of MKLP2, disturbs the organization of the anaphase spindle midzone. Using TIRF microscopy and purified fluorescent proteins, we here demonstrate that MKLP2 is a plus-end directed processive motor that can transport the CPC along microtubules in vitro. Live imaging of MKLP2::GFP and INCENP::GFP in early anaphase cells revealed that a fraction of both MKLP2 and INCENP displays directed movements towards the (equatorial) cortex. In line, inhibition of MKLP2 ATPase activity at the start of anaphase disturbed the localization of MKLP2 and CPC at the equatorial cortex. Our data suggest that MKLP2 is a processive microtubule-based motor that transports itself and the CPC to the equatorial cortex.
2020-06-10 | PXD014665 | Pride
Project description:Adenosine triphosphate (ATP) synthase is located on the inner membrane of mitochondria which generate ATP for various cellular processes. Our previous studies showed that ATP synthases existed on plasma membrane of several types of cancer cells, which was defined as ectopic ATP synthase. However, the trafficking pathway of ectopic ATP synthase is still unclear. To investigate how ATP synthase subunits are involved in the trafficking process, we performed shot-gun proteomic analysis to reveal the plasma membrane proteins of A549 lung cancer cells and identified four subunits of ATP synthases, ATP5A1, ATP5B, ATP5H, and MT-ATP6. With the results of gene set enrichment analysis, we inferred that ectopic ATP synthase subunits are assembled as complex consistently and then translocated to cell surface through the trafficking of mitochondria. On the other hand, microtubules are involved in many intracellular transport. During protein trafficking, mitochondria and vesicles are transported to their final destination along microtubules by motor proteins such as kinesins and dyneins. To explore the function of microtubules in the trafficking of ectopic ATP synthase, we treated MCF-7 breast and A549 lung cancer cells with nocodazole, a microtubule depolymerization agent, and found that the expression level of ectopic ATP synthase decreased using both immunocytochemistry and flow cytometry. Besides, since kinesin family member 5B (KIF5B) is an important motor proteins involved in the transport of mitochondria, we further performed the immunoprecipitation of KIF5B followed by liquid chromatography−tandem mass spectrometry (LC−MS/MS). The result revealed that one of the mitochondrial outer membrane proteins, dynamin-1 like proteins (Drp1), was able to associate to KIF5B. From our recent research results and other reports show that Drp1 is a mediator of mitochondrial dynamic through regulation of mitochondrial fission. Taken together, our findings suggested that Drp1-KIF5B complex-mediated mitochondrial trafficking along microtubules may play a critical role in the transport of ectopic ATP synthase
2023-05-10 | PXD006791 | Pride