Subgroup II PAK-mediated phosphorylation regulates Ran activity during mitosis.
Ran is an essential GTPase that controls nucleocytoplasmic transport, mitosis, and nuclear envelope formation. These functions are regulated by interaction of Ran with different partners, and by formation of a Ran-GTP gradient emanating from chromatin. Here, we identify a novel level of Ran regulation. We show that Ran is a substrate for p21-activated kinase 4 (PAK4) and that its phosphorylation on serine-135 increases during mitosis. The endogenous phosphorylated Ran and active PAK4 dynamically associate with different components of the microtubule spindle during mitotic progression. A GDP-bound Ran phosphomimetic mutant cannot undergo RCC1-mediated GDP/GTP exchange and cannot induce microtubule asters in mitotic Xenopus egg extracts. Conversely, phosphorylation of GTP-bound Ran facilitates aster nucleation. Finally, phosphorylation of Ran on serine-135 impedes its binding to RCC1 and RanGAP1. Our study suggests that PAK4-mediated phosphorylation of GDP- or GTP-bound Ran regulates the assembly of Ran-dependent complexes on the mitotic spindle.
Bompard
Guillaume
G
Universités Montpellier 2 et 1, IFR122, 34293 Montpellier, France.
Rabeharivelo
Gabriel
G
Frank
Marie
M
Cau
Julien
J
Delsert
Claude
C
Morin
Nathalie
N
D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism; D005260:Female; D006153:Guanosine Diphosphate / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D013379:Substrate Specificity; D014981:Xenopus / Q000235:genetics / Q000378:metabolism; D029867:Xenopus Proteins / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism; D054462:p21-Activated Kinases / Q000378:metabolism*; D020931:ran GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
D002843:Chromatin; D011993:Recombinant Fusion Proteins; D029867:Xenopus Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; C554182:PAK4 protein, Xenopus; D054462:p21-Activated Kinases; D020931:ran GTP-Binding Protein
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