Differential Modes of Nuclear Localization Signal (NLS) Recognition by Three Distinct Classes of NLS Receptors

• Published in: The Journal of biological chemistry Vol. 272; no. 42; pp. 26375 - 26381
• Main Authors: Miyamoto, Yoichi, Imamoto, Naoko, Sekimoto, Toshihiro, Tachibana, Taro, Seki, Takehiko, Tada, Shusuke, Enomoto, Takemi, Yoneda, Yoshihiro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.10.1997; American Society for Biochemistry and Molecular Biology
• Subjects: Adenosine Triphosphatases - metabolism; alpha Karyopherins; Amino Acid Sequence; Animals; Carrier Proteins - metabolism; Cattle; Cell Line; DNA Helicases - metabolism; HeLa Cells; Humans; Molecular Sequence Data; Nuclear Localization Signals; Nuclear Proteins - metabolism; Protein Binding; Recombinant Proteins - metabolism; RecQ Helicases
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.272.42.26375

The targeting of karyophilic proteins to nuclear pores is mediated via the formation of a nuclear pore-targeting complex, through the interaction of nuclear localization signal (NLS) with its NLS receptor. Recently, a novel human protein, Qip1, was identified from a yeast two-hybrid system with DNA helicase Q1. This study demonstrates that Qip1 is a novel third class of NLS receptor that efficiently recognizes the NLS of the helicase Q1. Moreover, the data obtained in this study show that the specific interaction between Qip1 and the NLS of the helicase Q1 requires its upstream sequence of the minimal essential NLS. By using purified recombinant proteins alone in the digitonin-permeabilized cell-free transport system, it was demonstrated that the two known human NLS receptors, Rch1 and NPI-1, are able to transport all the tested NLS substrates into the nucleus, while Qip1 most efficiently transports the helicase Q1-NLS substrates, which contain its upstream sequence in so far as we have examined the system. Furthermore, in HeLa cell crude cytosol, it was found that endogenous Rch1 binds to all the tested NLS substrates, while the binding of endogenous NPI-1 is restricted to only some NLSs, despite the fact that NPI-1 itself shows binding activity to a variety of NLSs. These results indicate that at least three structurally and functionally distinct NLS receptors exist in the human single cell population, and suggest that the nuclear import of karyophilic proteins may be controlled in a complex manner at the NLS recognition step by the existence of a variety of NLS receptors with various specificities to each NLS.