Transportin2 functions as importin and mediates nuclear import of HuR.
The RanGTP-binding nuclear transport receptors transportin1 (TRN1) and transportin2 (TRN2) are highly similar in sequence but are reported to function in nuclear import and export, respectively. Here we show that TRN2 possesses properties of a nuclear import receptor. TRN1/2 both interacted with a similar set of RNA-binding proteins in a RanGTP-sensitive manner. TRN2 bound RanGTP with high affinity, a feature of nuclear import receptors. As expected of an import complex, RanGTP also disrupted the interaction between TRN2 and HuR, an RNA-binding protein previously described as a TRN2 export substrate. The HuR nucleocytoplasmic shuttling signal, a sequence resembling the M9 nuclear import signal of hnRNP A1, was necessary and sufficient for TRN-mediated nuclear import of HuR in vitro. Finally, crosscompetition experiments demonstrated that HuR nucleocytoplasmic shuttling signal and M9 are imported along redundant pathways involving TRN1/2, substantiating the function of TRN2 in nuclear import.
Güttinger
Stephan
S
Swiss Federal Institute of Technology, Institute of Biochemistry, HPM F11.1, 8093 Zürich, Switzerland.
Mühlhäusser
Petra
P
Koller-Eichhorn
Roland
R
Brennecke
Julius
J
Kutay
Ulrike
U
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000954:Antigens, Surface / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D039102:Poly(A)-Binding Proteins / Q000378:metabolism; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011817:Rabbits; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism*
D000906:Antibodies; D000954:Antigens, Surface; D018076:DNA, Complementary; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D039102:Poly(A)-Binding Proteins; D016601:RNA-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; C503621:TNPO2 protein, human; D042461:Vascular Endothelial Growth Factor A; D028961:beta Karyopherins