Abstract
Signal-mediated protein transport through the nuclear pore complex is of considerable interest in the field of molecular pharmaceutics. Nuclear localization signals can be used to target genes/antisense delivery systems to the nucleus Studying nuclear export is useful in enhancing the expression and the efficiency of action, of these therapeutic agents. The mechanism of nuclear import has been well studied and most of the proteins participating in this mechanism have been identified. The subject of nuclear export is still in the initial stages and there is a considerable amount of uncertainty in this area. Two main export receptors identified so far are Exportin 1 (Crm1) and Calreticulin. Crm1 recognizes certain leucine-rich amino acid sequences in the proteins it exports called classical nuclear export signals. This paper describes a model system to study, identify, and establish these classical nuclear export signals using green fluorescent protein (GFP). Two putative export signals in the human progesterone receptor (PR) and the strongest nuclear export signal known (from mitogen activated protein kinase kinase [MAPKK]) were studied using this model system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bremner KH, Seymour LW, Pouton CW. Hamessing nuclear localization pathways for transgene delivery. Curr Opin Mol Ther. 2001;3:170–177.
Zanta MA, Belguise-Valladier P, Behr JP. Gene delivery: a single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus. Proc Natl Acad Sci U S A. 1999;96:91–96.
Ma H, Diamond SL. Nonviral gene therapy and its delivery systems. Curr Pharm Biotechnol. 2001;2:1–17.
Meunier L, Mayer R, Monsigny M, Roche AC. The nuclear export signal-dependent localization of oligonucleopeptides enhances the inhibition of the protein expression from a gene transcribed in cytosol. Nucleic Acids Res. 1999;27:2730–2736.
Stoffler D, Fahrenkrog B, Aebi U. The nuclear pore complex: from molecular architecture to functional dynamics. Curr Opin Cell Biol. 1999;11:391–401.
Kiseleva E, Goldberg MW, Cronshaw J, Allen TD. The nuclear pore complex: structure, function, and dynamics. Crit Rev Eukaryot Gene Expr. 2000;10:101–112.
Yoneda Y. Nucleocytoplasmic protein traffic and its significance to cell function. Genes Cells. 2000;5:777–787.
Macara IG. Transport into and out of the nucleus. Microbiol Mol Biol Rev. 2001;65:570–594.
Pruschy M, Ju Y, Spitz L, Carafoli E, Goldfarb DS. Facillitated nuclear transport of calmodulin in tissue culture cells. J Cell Biol. 1994;127:1527–1536.
Alberts DB, Lewis J, Raff M, Roberts K, Watson JD. Molecular Biology of the Cell. 2nd ed. New York, NY: Garland Publishing Inc; 1989.
Oxender DL, Quay S. Binding proteins and membrane transport. Ann NY Acad Sci. 1975;264:358–372.
Talcott B, Moore MS. Getting across the nuclear pore complex. Trends Cell Biol. 1999;9:312–318.
Htun H, Barsony J, Renyi I, Gould DL, Hager GL. Visualization of glucocorticoid receptor translocation and intranuclear organization in living cells with a green fluorescent protein chimera. Proc Natl Acad Sci U S A. 1996;93:4845–4850.
Kalderon D, Roberts BL, Richardson WD, Smith AE. A short amino acid sequence able to specify nuclear location. Cell. 1984;39:499–509.
Robbins J, Dilworth SM, Laskey RA, Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell. 1991;64:615–623.
Bogerd HP, Fridell RA, Benson RE, Hua J, Cullen BR. Protein sequence requirements for function of the human T-cell leukemia virus type 1 Rex nuclear export signal delineated by a novel in vivo randomization-selection assay. Mol Cell Biol. 1996;16:4207–4214.
Ikuta T, Eguchi H, Tachibana T, Yoneda Y, Kawajiri K. Nuclear localization and export signals of the human aryl hydrocarbon receptor. J Biol Chem. 1998;273:2895–2904.
Mattaj IW, Englmeier L. Nucleocytoplasmic transport: the soluble phase. Annu Rev Biochem. 1998;67:265–306.
Baumann CT, Lim CS, Hager GL. Simultaneous visualization of the yellow and green forms of the green fluorescent protein in living cells. J Histochem Cytochem. 1998;46:1073–1076.
Tsien RY. The green fluorescent protein. Annu Rev Biochem. 1998;67:509–544.
Patterson GH KS, Sharif WD, Kain SR, Piston DW. Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy. Biophys J. 1997;73:2782–2790.
McKenna NJ, Lanz RB, O'Malley BW. Nuclear receptor coregulators: cellular and molecular biology. Endocr Rev. 1999;20:321–344.
Defranco DB, Madan AP, Tang Y, Chandran UR, Xiao N, Yang J. Nucleocytoplasmic shuttling of steroid receptors. Vitam Horm. 1995;51:315–338.
Ostrowski MC R-FH, Wolford RG, Berard DS, Hager GL. Glucocorticoid regulation of transcription at an amplified, episomal promoter. Mol Cell Biol. 1983;11:2045–2057.
Henderson BR, Eleftheriou A. A comparison of the activity, sequence specificity, and CRM1-dependence of different nuclear export signals. Exp Cell Res. 2000;256:213–224.
Holaska JM, Black BE, Love DC, Hanover JA, Leszyk J, Paschal BM. Calreticulin is a receptor for nuclear export. J Cell Biol. 2001;152:127–140.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published: September 26, 2002
Rights and permissions
About this article
Cite this article
Kanwal, C., Li, H. & Lim, C.S. Model system to study classical nuclear export signals. AAPS J 4, 18 (2002). https://doi.org/10.1208/ps040318
Received:
Accepted:
Published:
DOI: https://doi.org/10.1208/ps040318