PMID- 10543952
OWN - NLM
STAT- MEDLINE
DCOM- 19991119
LR  - 20200331
IS  - 0022-2836 (Print)
IS  - 0022-2836 (Linking)
VI  - 293
IP  - 3
DP  - 1999 Oct 29
TI  - Interaction between NTF2 and xFxFG-containing nucleoporins is required to mediate
      nuclear import of RanGDP.
PG  - 579-93
AB  - Nuclear transport factor 2 (NTF2) is a small, homodimeric protein that binds to
      both RanGDP and xFxFG repeat-containing nucleoporins, such as yeast Nsp1p and
      vertebrate p62. NTF2 is required for efficient nuclear protein import and has
      been shown to mediate the nuclear import of RanGDP. We have used the crystal
      structures of rat NTF2 and its complex with RanGDP to design a mutant, W7A-NTF2, 
      in which the affinity for xFxFG-repeat nucleoporins is reduced while wild-type
      binding to RanGDP is retained. The 2.5 A resolution crystal structure of W7A-NTF2
      is virtually superimposable upon the wild-type protein structure, indicating that
      the mutation had not introduced a more general conformational change. Therefore, 
      our data suggest that the exposed side-chain of residue 7 is crucial to the
      interaction between NTF2 and xFxFG repeat-containing nucleoporins. Consistent
      with its reduced affinity for xFxFG nucleoporins, fluorescently labelled W7A-NTF2
      binds less strongly to the nuclear envelope of permeabilized cultured cells than 
      wild-type NTF2 and, when microinjected into Xenopus oocytes, colloidal gold
      coated with W7A-NTF2 binds less strongly to the central channel of nuclear pore
      complexes than wild-type NTF2-coated gold. Significantly, W7A-NTF2 only weakly
      stimulated the nuclear import of fluorescein-labelled RanGDP, providing direct
      evidence that an interaction between NTF2 and xFxFG repeat-containing
      nucleoporins is required to mediate the nuclear import of RanGDP.
CI  - Copyright 1999 Academic Press.
FAU - Bayliss, R
AU  - Bayliss R
AD  - MRC Laboratory of Molecular Biology, Hills Rd, Cambridge, CB2 2QH, England.
FAU - Ribbeck, K
AU  - Ribbeck K
FAU - Akin, D
AU  - Akin D
FAU - Kent, H M
AU  - Kent HM
FAU - Feldherr, C M
AU  - Feldherr CM
FAU - Gorlich, D
AU  - Gorlich D
FAU - Stewart, M
AU  - Stewart M
LA  - eng
SI  - PDB/1QMA
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Calcium-Binding Proteins)
RN  - 0 (Carrier Proteins)
RN  - 0 (Fungal Proteins)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (NSP1 protein, S cerevisiae)
RN  - 0 (Nuclear Pore Complex Proteins)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Nucleocytoplasmic Transport Proteins)
RN  - 0 (Nutf2 protein, mouse)
RN  - 0 (Nutf2 protein, rat)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 0 (nuclear pore protein p62)
RN  - EC 3.6.5.2 (ran GTP-Binding Protein)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Biological Transport
MH  - *Calcium-Binding Proteins
MH  - Carrier Proteins/*chemistry/genetics/isolation & purification/*metabolism
MH  - Cell Membrane Permeability
MH  - Crystallization
MH  - Crystallography, X-Ray
MH  - Fungal Proteins/chemistry/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Membrane Glycoproteins/chemistry/*metabolism
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Nuclear Envelope/metabolism/ultrastructure
MH  - Nuclear Pore Complex Proteins
MH  - Nuclear Proteins/*chemistry/genetics/isolation & purification/*metabolism
MH  - *Nucleocytoplasmic Transport Proteins
MH  - Oocytes/cytology/metabolism/ultrastructure
MH  - Protein Conformation
MH  - Rats
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/metabolism
MH  - *Repetitive Sequences, Amino Acid
MH  - *Saccharomyces cerevisiae Proteins
MH  - Solubility
MH  - Xenopus laevis
MH  - ran GTP-Binding Protein/*metabolism
EDAT- 1999/11/02 00:00
MHDA- 1999/11/02 00:01
CRDT- 1999/11/02 00:00
PHST- 1999/11/02 00:00 [pubmed]
PHST- 1999/11/02 00:01 [medline]
PHST- 1999/11/02 00:00 [entrez]
AID - 10.1006/jmbi.1999.3166 [doi]
AID - S0022-2836(99)93166-2 [pii]
PST - ppublish
SO  - J Mol Biol. 1999 Oct 29;293(3):579-93. doi: 10.1006/jmbi.1999.3166.