A karyophilic protein forms a stable complex with cytoplasmic components prior to nuclear pore binding

Abstract

Targeting of karyophilic proteins to nuclear pores is known to require several cytoplasmic factors, including the nuclear location signal-binding protein. Using a digitonin-permeabilized cell-free transport assay, we have obtained a cytoplasmic fraction containing factors that specifically bind to karyophilic protein and support the nuclear binding step of the transport. Components in this fraction form a stable complex with the karyophile through interaction with nuclear location signal. Since this complex shows nuclear pore binding activity prior to nuclear entry in the absence of other cytosolic factors, we call it nuclear pore-targeting complex. It consists of karyophilic protein and four proteins of 54, 56, 66, and 90 kDa. In our reconstitution experiments, a complex with 54 and 90 kDa proteins is capable of targeting karyophiles to the nuclear pores.