RAN/TC4 mutants identify a common requirement for snRNP and protein import into the nucleus.
Kinetic competition experiments have demonstrated that at least some factors required for the nuclear import of proteins and U snRNPs are distinct. Both import processes require energy, and in the case of protein import, the energy requirement is known to be at least partly met by GTP hydrolysis by the Ran GTPase. We have compared the effects of nonhydrolyzable GTP analogues and two mutant Ran proteins on the nuclear import of proteins and U snRNPs in vitro. The mutant Ran proteins have different defects; Q69L (glutamine 69 changed to leucine) is defective in GTP hydrolysis while T24N (threonine 24 changed to asparagine) is defective in binding GTP. Both protein and snRNP import are sensitive either to the presence of the two mutant Ran proteins, which act as dominant negative inhibitors of nuclear import, or to incubation with nonhydrolyzable GTP analogues. This demonstrates that there is a requirement for a GTPase activity for the import of U snRNPs, as well as proteins, into the nucleus. The dominant negative effects of the two mutant Ran proteins indicate that the pathways of protein and snRNP import share at lease one common component.
Palacios
I
I
Gene Expression Programme, European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
Weis
K
K
Klebe
C
C
Mattaj
I W
IW
Dingwall
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D009685:Nuclear Envelope / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D012314:RNA Cap Analogs / Q000378:metabolism / Q000494:pharmacology; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism*; D014981:Xenopus; D020931:ran GTP-Binding Protein
D009687:Nuclear Proteins; D012314:RNA Cap Analogs; D017411:Ribonucleoproteins, Small Nuclear; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D020931:ran GTP-Binding Protein