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1.
Elife ; 82019 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-31808746

RESUMO

Microtubules segregate chromosomes by attaching to macromolecular kinetochores. Only microtubule-end attached kinetochores can be pulled apart; how these end-on attachments are selectively recognised and stabilised is not known. Using the kinetochore and microtubule-associated protein, Astrin, as a molecular probe, we show that end-on attachments are rapidly stabilised by spatially-restricted delivery of PP1 near the C-terminus of Ndc80, a core kinetochore-microtubule linker. PP1 is delivered by the evolutionarily conserved tail of Astrin and this promotes Astrin's own enrichment creating a highly-responsive positive feedback, independent of biorientation. Abrogating Astrin:PP1-delivery disrupts attachment stability, which is not rescued by inhibiting Aurora-B, an attachment destabiliser, but is reversed by artificially tethering PP1 near the C-terminus of Ndc80. Constitutive Astrin:PP1-delivery disrupts chromosome congression and segregation, revealing a dynamic mechanism for stabilising attachments. Thus, Astrin-PP1 mediates a dynamic 'lock' that selectively and rapidly stabilises end-on attachments, independent of biorientation, and ensures proper chromosome segregation.


Assuntos
Azul Alciano/metabolismo , Segregação de Cromossomos , Cinetocoros/metabolismo , Microtúbulos/metabolismo , Fenazinas/metabolismo , Fenotiazinas/metabolismo , Receptores de Neuropeptídeo Y/metabolismo , Resorcinóis/metabolismo , Azul Alciano/química , Aurora Quinase B , Proteínas Cromossômicas não Histona , Proteínas do Citoesqueleto/metabolismo , Células HeLa , Humanos , Cinetocoros/química , Metáfase , Proteínas Associadas aos Microtúbulos/metabolismo , Simulação de Acoplamento Molecular , Fenazinas/química , Fenotiazinas/química , Conformação Proteica , Domínios e Motivos de Interação entre Proteínas , Receptores de Neuropeptídeo Y/química , Receptores de Neuropeptídeo Y/genética , Resorcinóis/química
2.
Open Biol ; 9(6): 180263, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31238822

RESUMO

Tissue maintenance and development requires a directed plane of cell division. While it is clear that the division plane can be determined by retraction fibres that guide spindle movements, the precise molecular components of retraction fibres that control spindle movements remain unclear. We report MARK2/Par1b kinase as a novel component of actin-rich retraction fibres. A kinase-dead mutant of MARK2 reveals MARK2's ability to monitor subcellular actin status during interphase. During mitosis, MARK2's localization at actin-rich retraction fibres, but not the rest of the cortical membrane or centrosome, is dependent on its activity, highlighting a specialized spatial regulation of MARK2. By subtly perturbing the actin cytoskeleton, we reveal MARK2's role in correcting mitotic spindle off-centring induced by actin disassembly. We propose that MARK2 provides a molecular framework to integrate cortical signals and cytoskeletal changes in mitosis and interphase.


Assuntos
Actinas/metabolismo , Centrossomo/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Fuso Acromático/metabolismo , Células HeLa , Humanos , Mitose , Mutação , Proteínas Serina-Treonina Quinases/genética
3.
J Cell Biol ; 217(9): 3057-3070, 2018 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-29941476

RESUMO

The plane of cell division is defined by the final position of the mitotic spindle. The spindle is pulled and rotated to the correct position by cortical dynein. However, it is unclear how the spindle's rotational center is maintained and what the consequences of an equatorially off centered spindle are in human cells. We analyzed spindle movements in 100s of cells exposed to protein depletions or drug treatments and uncovered a novel role for MARK2 in maintaining the spindle at the cell's geometric center. Following MARK2 depletion, spindles glide along the cell cortex, leading to a failure in identifying the correct division plane. Surprisingly, spindle off centering in MARK2-depleted cells is not caused by excessive pull by dynein. We show that MARK2 modulates mitotic microtubule growth and length and that codepleting mitotic centromere-associated protein (MCAK), a microtubule destabilizer, rescues spindle off centering in MARK2-depleted cells. Thus, we provide the first insight into a spindle-centering mechanism needed for proper spindle rotation and, in turn, the correct division plane in human cells.


Assuntos
Mitose/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Fuso Acromático/metabolismo , Linhagem Celular Tumoral , Dineínas/metabolismo , Células HeLa , Humanos , Microtúbulos/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética
4.
J Food Sci Technol ; 52(4): 2272-9, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25829609

RESUMO

The present study investigates the antimicrobial and preservative potentials of turmeric extracts for food industry. Turmeric extracts prepared in n-hexane, water, chloroform and ethanol were applied to meals as preservatives and antibacterial agent. The samples were assessed microbiologically (total bacterial, total fungal and total coliform counts) and organoleptically (color, odor, taste) at day zero and after 15 days intervals. Meals autoclaved for shorter time (5 min) and treated with combination of 1 % or 2 % turmeric extract preserved for longer period. These results were comparable with samples autoclaved for longer period (15 min) with out turmeric extract. The antibacterial activities of different turmeric extracts were also tested against Escherichia coli, Staphylococcus aureus, Salmonella typhi and Candida albicans by disc diffusion method. Water extracted samples of turmeric stored at room temperature inhibited the growth of Escherichia coli and Salmonella typhi while aqueous extract autoclaved at 121 °C for 30 min reduced the growth of Escherichia coli and Staphylococcus aureus. Methanol extracted samples stored at room temperature or autoclaved at 121 °C was effective to control the growth of all microbes under study. Chloroform and n-hexane extracts (stored at room temperature) showed weak activity against all tested microbes.

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