The Journal of Cell Biology, Vol 107, 1279-1287, Copyright © 1988 by The Rockefeller University Press

ARTICLES

The effects of variations in the number and sequence of targeting signals on nuclear uptake

SI Dworetzky, RE Lanford and CM Feldherr
Department of Anatomy and Cell Biology, University of Florida, College of Medicine, Gainesville 32610.

To determine if the number of targeting signals affects the transport of proteins into the nucleus, Xenopus oocytes were injected with colloidal gold particles, ranging in diameter from 20 to 280 A, that were coated with BSA cross-linked with synthetic peptides containing the SV-40 large T-antigen nuclear transport signal. Three BSA conjugate preparations were used; they had an average of 5, 8, and 11 signals per molecule of carrier protein. In addition, large T-antigen, which contains one signal per monomer, was used as a coating agent. The cells were fixed at various times after injection and subsequently analyzed by electron microscopy. Gold particles coated with proteins containing the SV-40 signal entered the nucleus through central channels located within the nuclear pores. Analysis of the intracellular distribution and size of the tracers that entered the nucleus indicated that the number of signals per molecule affect both the relative uptake of particles and the functional size of the channels available for translocation. In control experiments, gold particles coated with BSA or BSA conjugated with inactive peptides similar to the SV-40 transport signal were virtually excluded from the nucleus. Gold particles coated with nucleoplasmin, an endogenous karyophilic protein that contains five targeting signals per molecule, was transported through the nuclear pores more effectively than any of the BSA-peptide conjugates. Based on a correlation between the peri-envelope density of gold particles and their relative uptake, it is suggested that the differences in the activity of the two targeting signals is related to their binding affinity for envelope receptors. It was also determined, by performing coinjection experiments, that individual pores are capable of recognizing and transporting proteins that contain different nuclear targeting signals.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Naim, B., Brumfeld, V., Kapon, R., Kiss, V., Nevo, R., Reich, Z. (2007). Passive and Facilitated Transport in Nuclear Pore Complexes Is Largely Uncoupled. J. Biol. Chem. 282: 3881-3888 [Abstract] [Full Text]  
• Khalil, I. A., Kogure, K., Akita, H., Harashima, H. (2006). Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery.. Pharmacol. Rev. 58: 32-45 [Abstract] [Full Text]  
• Irwin, B., Aye, M., Baldi, P., Beliakova-Bethell, N., Cheng, H., Dou, Y., Liou, W., Sandmeyer, S. (2005). Retroviruses and yeast retrotransposons use overlapping sets of host genes. Genome Res 15: 641-654 [Abstract] [Full Text]  
• Liu, G., Li, D., Pasumarthy, M. K., Kowalczyk, T. H., Gedeon, C. R., Hyatt, S. L., Payne, J. M., Miller, T. J., Brunovskis, P., Fink, T. L., Muhammad, O., Moen, R. C., Hanson, R. W., Cooper, M. J. (2003). Nanoparticles of Compacted DNA Transfect Postmitotic Cells. J. Biol. Chem. 278: 32578-32586 [Abstract] [Full Text]  
• Quadrini, K. J., Bieker, J. J. (2002). Kruppel-like Zinc Fingers Bind to Nuclear Import Proteins and Are Required for Efficient Nuclear Localization of Erythroid Kruppel-like Factor. J. Biol. Chem. 277: 32243-32252 [Abstract] [Full Text]  
• Nakanishi, A., Shum, D., Morioka, H., Otsuka, E., Kasamatsu, H. (2002). Interaction of the Vp3 Nuclear Localization Signal with the Importin {alpha}2/{beta} Heterodimer Directs Nuclear Entry of Infecting Simian Virus 40. J. Virol. 76: 9368-9377 [Abstract] [Full Text]  
• Feldherr, C., Akin, D., Littlewood, T., Stewart, M. (2002). The molecular mechanism of translocation through the nuclear pore complex is highly conserved. J. Cell Sci. 115: 2997-3005 [Abstract] [Full Text]  
• Feldherr, C. M., Akin, D., Cohen, R. J. (2002). Regulation of functional nuclear pore size in fibroblasts. J. Cell Sci. 114: 4621-4627 [Abstract] [Full Text]  
• Vihinen-Ranta, M., Wang, D., Weichert, W. S., Parrish, C. R. (2002). The VP1 N-Terminal Sequence of Canine Parvovirus Affects Nuclear Transport of Capsids and Efficient Cell Infection. J. Virol. 76: 1884-1891 [Abstract] [Full Text]  
• Salman, H., Zbaida, D., Rabin, Y., Chatenay, D., Elbaum, M. (2001). Kinetics and mechanism of DNA uptake into the cell nucleus. Proc. Natl. Acad. Sci. USA 10.1073/pnas.121067698v1 [Abstract] [Full Text]  
• Lechardeur, D., Drzymala, L., Sharma, M., Zylka, D., Kinach, R., Pacia, J., Hicks, C., Usmani, N., Rommens, J. M., Lukacs, G. L. (2000). Determinants of the Nuclear Localization of the Heterodimeric DNA Fragmentation Factor (Icad/Cad). JCB 150: 321-334 [Abstract] [Full Text]  
• Vihinen-Ranta, M., Yuan, W., Parrish, C. R. (2000). Cytoplasmic Trafficking of the Canine Parvovirus Capsid and Its Role in Infection and Nuclear Transport. J. Virol. 74: 4853-4859 [Abstract] [Full Text]  
• Sherman, D. L., Brophy, P. J. (2000). A Tripartite Nuclear Localization Signal in the PDZ-domain Protein L-periaxin. J. Biol. Chem. 275: 4537-4540 [Abstract] [Full Text]  
• Wilson, G. L., Dean, B. S., Wang, G., Dean, D. A. (1999). Nuclear Import of Plasmid DNA in Digitonin-permeabilized Cells Requires Both Cytoplasmic Factors and Specific DNA Sequences. J. Biol. Chem. 274: 22025-22032 [Abstract] [Full Text]  
• Bodoor, K, Shaikh, S, Salina, D, Raharjo, W., Bastos, R, Lohka, M, Burke, B (1999). Sequential recruitment of NPC proteins to the nuclear periphery at the end of mitosis. J. Cell Sci. 112: 2253-2264 [Abstract]  
• Ludtke, J., Zhang, G, Sebestyen, M., Wolff, J. (1999). A nuclear localization signal can enhance both the nuclear transport and expression of 1 kb DNA. J. Cell Sci. 112: 2033-2041 [Abstract]  
• Feldherr, C., Akin, D (1999). Signal-mediated nuclear transport in the amoeba. J. Cell Sci. 112: 2043-2048 [Abstract]  
• Plafker, S. M., Gibson, W. (1998). Cytomegalovirus Assembly Protein Precursor and Proteinase Precursor Contain Two Nuclear Localization Signals That Mediate Their Own Nuclear Translocation and That of the Major Capsid Protein. J. Virol. 72: 7722-7732 [Abstract] [Full Text]  
• Shah, S., Tugendreich, S., Forbes, D. (1998). Major Binding Sites for the Nuclear Import Receptor Are the Internal Nucleoporin Nup153 and the Adjacent Nuclear Filament Protein Tpr. JCB 141: 31-49 [Abstract] [Full Text]  
• Feldherr, C, Akin, D, Moore, M. (1998). The nuclear import factor p10 regulates the functional size of the nuclear pore complex during oogenesis. J. Cell Sci. 111: 1889-1896 [Abstract]  
• Shields, J. M., Yang, V. W. (1997). Two Potent Nuclear Localization Signals in the Gut-enriched Kruppel-like Factor Define a Subfamily of Closely Related Kruppel Proteins. J. Biol. Chem. 272: 18504-18507 [Abstract] [Full Text]  
• Feldherr, C., Akin, D (1997). The location of the transport gate in the nuclear pore complex. J. Cell Sci. 110: 3065-3070 [Abstract]  
• Rout, M. P., Aitchison, J. D. (2001). The Nuclear Pore Complex as a Transport Machine. J. Biol. Chem. 276: 16593-16596 [Full Text]  
• Salman, H., Zbaida, D., Rabin, Y., Chatenay, D., Elbaum, M. (2001). Kinetics and mechanism of DNA uptake into the cell nucleus. Proc. Natl. Acad. Sci. USA 98: 7247-7252 [Abstract] [Full Text]  
• Pante, N., Kann, M. (2002). Nuclear Pore Complex Is Able to Transport Macromolecules with Diameters of ~39 nm. Mol. Biol. Cell 13: 425-434 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents