Kap95p binding induces the switch loops of RanGDP to adopt the GTP-bound conformation: implications for nuclear import complex assembly dynamics

Jade Forwood, Thierry Lonhienne, Mary Marfori, Robin Gautier, Weining Meng, Gregor Guncar, Sai M Liu, Murray Stewart, Bernard Carroll, Bostjan Kobe

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Abstract

The asymmetric distribution of the nucleotide-bound state of Ran across the nuclear envelope is crucial for determining the directionality of nuclear transport. In the nucleus, Ran is primarily in the guanosine 5'-triphosphate (GTP)-bound state, whereas in the cytoplasm, Ran is primarily guanosine 5'-diphosphate (GDP)-bound. Conformational changes within the Ran switch I and switch II loops are thought to modulate its affinity for importin-beta. Here, we show that RanGDP and importin-beta form a stable complex with a micromolar dissociation constant. This complex can be dissociated by importin-beta binding partners such as importin-alpha. Surprisingly, the crystal structure of the Kap95p-RanGDP complex shows that Kap95p induces the switch I and II regions of RanGDP to adopt a conformation that resembles that of the GTP-bound form. The structure of the complex provides insights into the structural basis for the gradation of affinities regulating nuclear protein transport.
Original languageEnglish
Pages (from-to)772-782
Number of pages11
JournalJournal of Molecular Biology
Volume383
Issue number4
DOIs
Publication statusPublished - 2008

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beta Karyopherins
Cell Nucleus Active Transport
Guanosine Triphosphate
alpha Karyopherins
Guanosine
Diphosphates
Nuclear Envelope
Protein Transport
Nuclear Proteins
Cytoplasm
Nucleotides

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Forwood, Jade ; Lonhienne, Thierry ; Marfori, Mary ; Gautier, Robin ; Meng, Weining ; Guncar, Gregor ; Liu, Sai M ; Stewart, Murray ; Carroll, Bernard ; Kobe, Bostjan. / Kap95p binding induces the switch loops of RanGDP to adopt the GTP-bound conformation : implications for nuclear import complex assembly dynamics. In: Journal of Molecular Biology. 2008 ; Vol. 383, No. 4. pp. 772-782.
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abstract = "The asymmetric distribution of the nucleotide-bound state of Ran across the nuclear envelope is crucial for determining the directionality of nuclear transport. In the nucleus, Ran is primarily in the guanosine 5'-triphosphate (GTP)-bound state, whereas in the cytoplasm, Ran is primarily guanosine 5'-diphosphate (GDP)-bound. Conformational changes within the Ran switch I and switch II loops are thought to modulate its affinity for importin-beta. Here, we show that RanGDP and importin-beta form a stable complex with a micromolar dissociation constant. This complex can be dissociated by importin-beta binding partners such as importin-alpha. Surprisingly, the crystal structure of the Kap95p-RanGDP complex shows that Kap95p induces the switch I and II regions of RanGDP to adopt a conformation that resembles that of the GTP-bound form. The structure of the complex provides insights into the structural basis for the gradation of affinities regulating nuclear protein transport.",
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Kap95p binding induces the switch loops of RanGDP to adopt the GTP-bound conformation : implications for nuclear import complex assembly dynamics. / Forwood, Jade; Lonhienne, Thierry; Marfori, Mary; Gautier, Robin; Meng, Weining; Guncar, Gregor; Liu, Sai M; Stewart, Murray; Carroll, Bernard; Kobe, Bostjan.

In: Journal of Molecular Biology, Vol. 383, No. 4, 2008, p. 772-782.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Kap95p binding induces the switch loops of RanGDP to adopt the GTP-bound conformation

T2 - implications for nuclear import complex assembly dynamics

AU - Forwood, Jade

AU - Lonhienne, Thierry

AU - Marfori, Mary

AU - Gautier, Robin

AU - Meng, Weining

AU - Guncar, Gregor

AU - Liu, Sai M

AU - Stewart, Murray

AU - Carroll, Bernard

AU - Kobe, Bostjan

N1 - Imported on 12 Apr 2017 - DigiTool details were: Journal title (773t) = Journal of Molecular Biology. ISSNs: 0022-2836;

PY - 2008

Y1 - 2008

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AB - The asymmetric distribution of the nucleotide-bound state of Ran across the nuclear envelope is crucial for determining the directionality of nuclear transport. In the nucleus, Ran is primarily in the guanosine 5'-triphosphate (GTP)-bound state, whereas in the cytoplasm, Ran is primarily guanosine 5'-diphosphate (GDP)-bound. Conformational changes within the Ran switch I and switch II loops are thought to modulate its affinity for importin-beta. Here, we show that RanGDP and importin-beta form a stable complex with a micromolar dissociation constant. This complex can be dissociated by importin-beta binding partners such as importin-alpha. Surprisingly, the crystal structure of the Kap95p-RanGDP complex shows that Kap95p induces the switch I and II regions of RanGDP to adopt a conformation that resembles that of the GTP-bound form. The structure of the complex provides insights into the structural basis for the gradation of affinities regulating nuclear protein transport.

KW - Open access version available

KW - Importin

KW - Karyopherin

KW - Nucleocytoplasmic transport

KW - Protein structure

KW - Ran

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DO - 10.1016/j.jmb.2008.07.090

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