title: Expanding the definition of the classical bipartite nuclear localization signal.
abstract: Nuclear localization signals (NLSs) are amino acid sequences that target cargo proteins into the nucleus. Rigorous characterization of NLS motifs is essential to understanding and predicting pathways for nuclear import. The best-characterized NLS is the classical NLS (cNLS), which is recognized by the cNLS receptor, importin-alpha. cNLSs are conventionally defined as having one (monopartite) or two clusters of basic amino acids separated by a 9-12 aa linker (bipartite). Motivated by the finding that Ty1 integrase, which contains an unconventional putative bipartite cNLS with a 29 aa linker, exploits the classical nuclear import machinery, we assessed the functional boundaries for linker length within a bipartite cNLS. We confirmed that the integrase cNLS is a bona fide bipartite cNLS, then carried out a systematic analysis of linker length in an obligate bipartite cNLS cargo, which revealed that some linkers longer than conventionally defined can function in nuclear import. Linker function is dependent on the sequence and likely the inherent flexibility of the linker. Subsequently, we interrogated the Saccharomyces cerevisiae proteome to identify cellular proteins containing putative long bipartite cNLSs. We experimentally confirmed that Rrp4 contains a bipartite cNLS with a 25 aa linker. Our studies show that the traditional definition of bipartite cNLSs is too restrictive and linker length can vary depending on amino acid composition.
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, 1510 Clifton Road NE, Atlanta, GA 30322, USA.
authors.forename: Allison
authors.initials: A
authors.lastname: Lange
authors.forename: Laura M
authors.initials: LM
authors.lastname: McLane
authors.forename: Ryan E
authors.initials: RE
authors.lastname: Mills
authors.forename: Scott E
authors.initials: SE
authors.lastname: Devine
authors.forename: Anita H
authors.initials: AH
authors.lastname: Corbett
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D019426:Integrases / Q000378:metabolism; D028884:Karyopherins / Q000378:metabolism; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000378:metabolism*; D020543:Proteome; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D028884:Karyopherins; D019913:Nuclear Localization Signals; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins; D019426:Integrases