RESUMO
Coordination between the microtubule and actin networks is essential for cell motility, neuronal growth cone guidance, and wound healing. Members of the CLASP (cytoplasmic linker-associated protein) family of proteins have been implicated in the cytoskeletal cross-talk between microtubules and actin networks; however, the molecular mechanisms underlying the role of CLASP in cytoskeletal coordination are unclear. Here, we investigate CLASP2α's cross-linking function with microtubules and F-actin. Our results demonstrate that CLASP2α cross-links F-actin to the microtubule lattice in vitro. We find that the cross-linking ability is retained by L-TOG2-S, a minimal construct containing the TOG2 domain and serine-arginine-rich region of CLASP2α. Furthermore, CLASP2α promotes the accumulation of multiple actin filaments along the microtubule, supporting up to 11 F-actin landing events on a single microtubule lattice region. CLASP2α also facilitates the dynamic organization of polymerizing actin filaments templated by the microtubule network, with F-actin forming bridges between individual microtubules. Finally, we find that depletion of CLASPs in vascular smooth muscle cells results in disorganized actin fibers and reduced coalignment of actin fibers with microtubules, suggesting that CLASP and microtubules contribute to higher-order actin structures. Taken together, our results indicate that CLASP2α can directly cross-link F-actin to microtubules and that this microtubule-CLASP-actin interaction may influence overall cytoskeletal organization in cells.
Assuntos
Citoesqueleto de Actina , Actinas , Microtúbulos , Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/ultraestrutura , Actinas/metabolismo , Citoesqueleto/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Microtúbulos/ultraestrutura , Ligação Proteica , HumanosRESUMO
Microtubules are cytoskeletal polymers that dynamically remodel to perform essential cellular functions. Individual microtubules alternate between phases of growth and shrinkage via sudden transitions called catastrophe and rescue, driven by losing and regaining a stabilizing cap at the dynamic microtubule end. New in vitro studies now show that a conserved family of CLASP proteins specifically modulate microtubule catastrophe and rescue transitions. Further, recent cryo-electron microscopy approaches have elucidated new structural features of the stabilizing cap. Together, these new advances provide a clearer view on the complexity of the microtubule end and its regulation.
Assuntos
Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Animais , Microscopia Crioeletrônica , Humanos , Microtúbulos/ultraestrutura , Nucleotídeos/metabolismo , Tubulina (Proteína)/metabolismoRESUMO
Olive is one of the most important cultivated Mediterranean plants. In order to determine the differences in frost resistance of two, two-year-old olive cultivars (Olea europaea cv. Leccino and cv. Oblica) growing on different types of nutrient substrates (soil and coconut fibres), the trees were exposed to low temperature (-5 °C) in the dark. It was shown that low temperature caused an increase in H2O2 concentration, level of lipid peroxidation and carbonyl protein content in both cultivars and on both nutrient substrates, respectively. The CAT and APX activities significantly varied depending on the cultivar, the nutrient substrate type and the time of exposure to low temperature. Cv. Oblica and cv. Leccino growing on coconut fibres showed a better antioxidative response to low temperature probably due to the higher nitrogen and phosphorus concentration established in this type of nutrient substrate. That positive antioxidative response determined on coconut fibres was more pronounced in leaves of cv. Leccino.