RESUMO
Microtubule (MT)-associated protein 7 (MAP7) is a required cofactor for kinesin-1-driven transport of intracellular cargoes. Using cryo-electron microscopy and single-molecule imaging, we investigated how MAP7 binds MTs and facilitates kinesin-1 motility. The MT-binding domain (MTBD) of MAP7 bound MTs as an extended α helix between the protofilament ridge and the site of lateral contact. Unexpectedly, the MTBD partially overlapped with the binding site of kinesin-1 and inhibited its motility. However, by tethering kinesin-1 to the MT, the projection domain of MAP7 prevented dissociation of the motor and facilitated its binding to available neighboring sites. The inhibitory effect of the MTBD dominated as MTs became saturated with MAP7. Our results reveal biphasic regulation of kinesin-1 by MAP7 in the context of their competitive binding to MTs.
Assuntos
Cinesinas , Proteínas Associadas aos Microtúbulos , Microtúbulos , Humanos , Sítios de Ligação , Ligação Competitiva , Microscopia Crioeletrônica , Dineínas/química , Dineínas/metabolismo , Cinesinas/química , Cinesinas/metabolismo , Proteínas Associadas aos Microtúbulos/química , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Modelos Biológicos , Modelos Moleculares , Ligação Proteica , Conformação Proteica em alfa-Hélice , Domínios Proteicos , Tubulina (Proteína)/química , Tubulina (Proteína)/metabolismoRESUMO
A key process in the life of any multicellular organism is its development from a single egg into a full grown adult. The first step in this process often consists of forming a tissue layer out of randomly placed cells on the surface of the egg. We present a model for generating such a tissue, based on mechanical interactions between the cells, and find that the resulting cellular pattern corresponds to the Voronoi tessellation of the nuclei of the cells. Experimentally, we obtain the same result in both fruit flies and flour beetles, with a distribution of cell shapes that matches that of the model, without any adjustable parameters. Finally, we show that this pattern is broken when the cells grow at different rates.