Domains of Crm1 involved in the formation of the Crm1, RanGTP, and leucine-rich nuclear export sequences trimeric complex.
Nuclear export of proteins containing a leucine-rich nuclear export sequence (NES) is mediated by a specific NES receptor known as Crm1. This protein, which is related to the karyopherin beta family, interacts directly with NES in a RanGTP-dependent manner. To characterize the domains of Crm1 involved in formation of the trimeric Crm1-NES-RanGTP complex, N- and C-terminal deletion mutants of Crm1 were generated and their ability to bind NES and RanGTP in vitro was analyzed. Our results indicate that two regions of Crm1 are required for the formation of the trimeric Crm1-NES-RanGTP complex, the N-terminal domain of Crm1 and the central domain of the receptor, starting after residue 160 with an essential region between 566 and 720. The N-terminal domain is homologous to the RanGTP-binding domain of karyopherin beta and therefore is likely involved in the interaction with RanGTP. Consequently, the central domain likely corresponds to the NES-binding site of Crm1.
Ossareh-Nazari
B
B
Laboratoire de Transport Nucleocytoplasmique, Institut Curie-CNRS UMR144, 26 rue d'Ulm, Paris Cedex 05, 75248, France.
Dargemont
C
C
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D028884:Karyopherins; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020931:ran GTP-Binding Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000097606:Exportin 1 Protein
D002352:Carrier Proteins; D028884:Karyopherins; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D020931:ran GTP-Binding Protein; D000097606:Exportin 1 Protein