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The nucleoporin Nup50 activates the Ran guanyl-nucleotide exchange factor RCC1 to promote NPC assembly at the end of mitosis

ABSTRACT: During mitotic exit, thousands of nuclear pore complexes (NPCs) assemble concomitant with the nuclear envelope to build a transport-competent nucleus. Here, we show that Nup50 plays a crucial role in NPC assembly independent of its well-established function in nuclear transport. RNAi-mediated downregulation in cells or immunodepletion of Nup50 protein in Xenopus egg extracts interferes with NPC assembly. We define a conserved central region of 46 residues in Nup50 that is crucial for Nup153 and MEL28/ELYS binding, and for NPC interaction. Surprisingly, neither NPC interaction nor binding of Nup50 to importin /, the GTPase Ran, or chromatin is crucial for its function in the assembly process. Instead, an N-terminal fragment of Nup50 can stimulate the Ran GTPase guanyl-nucleotide exchange factor RCC1 and NPC assembly, indicating that Nup50 acts via the Ran system in NPC reformation at the end of mitosis. In support of this conclusion, Nup50 mutants defective in RCC1 binding and stimulation cannot replace the wild-type protein in in vitro NPC assembly assays, while excess RCC1 can compensate the loss of Nup50.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Christian Preisinger

LAB HEAD: Wolfram Antonin

PROVIDER: PXD028701 | Pride | 2021-11-03

REPOSITORIES: Pride

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			Action	DRS
	MQ_output.zip	Other
	OR2_2020_10_31_WoAnt_IP_NUP50_1_1.raw	Raw
	OR2_2020_10_31_WoAnt_IP_ctrl1_1.raw	Raw
	OR2_2021_02_01_WoAnt_IP_NUP50_2.raw	Raw
	OR2_2021_02_01_WoAnt_IP_NUP50_3.raw	Raw

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