This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zacksenhaus, E Articles by Gallie, B L Search for Related Content PubMed PubMed Citation Articles by Zacksenhaus, E Articles by Gallie, B L

A bipartite nuclear localization signal in the retinoblastoma gene product and its importance for biological activity.

Division of Immunology and Cancer Research, Hospital for Sick Children, Toronto, Ontario, Canada.

ABSTRACT

The retinoblastoma gene product, p110RB1, appears to regulate cell growth by modulating the activities of nuclear transcription factors. The elements that specify the transport of p110RB1 into the nucleus have not yet been explored. We now report the identification of a basic region, KRSAEGGNPPKPLKKLR, in the C terminus of p110RB1, which has sequence similarity to known bipartite nuclear localization signals (NLSs). A two-amino-acid mutation introduced into this putative NLS [to give mutant NLS(NQ)] or deletion of the entire NLS (delta NLS) abrogated exclusive nuclear localization, yielding proteins which were distributed either equally throughout the cell or predominantly in the cytoplasm. A mutant protein [NLS(NQ)/delta 22] containing both the mutated NLS and a deletion of exon 22, previously shown to disrupt the interaction of p110RB1 with several cellular transcription factors and oncoproteins, accumulated only in the cytoplasm. When fused to the C terminus of Escherichia coli beta-galactosidase, the RB1 NLS directed this protein to the nucleus, indicating that the motif is not only necessary but also sufficient for nuclear transport. Neither NLS(NQ) nor delta NLS was hyperphosphorylated in vivo, but both retained their abilities to interact, in vitro, with simian virus 40 large T antigen, adenovirus E1a, and the cellular transcription factor E2F. When transfected at multiple copy number, the NLS mutant alleles displayed reduced biological activity, measured by inhibition of growth of the osteogenic sarcoma cell line Saos-2, which has no wild-type RB1. Naturally occurring mutations and deletions in exon 25 of RB1 which disrupt the NLS may lead to partial or complete inactivation of p110RB1 and may be responsible for some retinoblastoma and other tumors.

This article has been cited by other articles:

• Nguyen, N. L., Loveland, A. N., Gibson, W. (2008). Nuclear Localization Sequences in Cytomegalovirus Capsid Assembly Proteins (UL80 Proteins) Are Required for Virus Production: Inactivating NLS1, NLS2, or Both Affects Replication to Strikingly Different Extents. J. Virol. 82: 5381-5389 [Abstract] [Full Text]  
• Li, C., Wu, R.-C., Amazit, L., Tsai, S. Y., Tsai, M.-J., O'Malley, B. W. (2007). Specific Amino Acid Residues in the Basic Helix-Loop-Helix Domain of SRC-3 Are Essential for Its Nuclear Localization and Proteasome-Dependent Turnover. Mol. Cell. Biol. 27: 1296-1308 [Abstract] [Full Text]  
• Jiao, W., Datta, J., Lin, H.-M., Dundr, M., Rane, S. G. (2006). Nucleocytoplasmic Shuttling of the Retinoblastoma Tumor Suppressor Protein via Cdk Phosphorylation-dependent Nuclear Export. J. Biol. Chem. 281: 38098-38108 [Abstract] [Full Text]  
• St-Pierre, B., Liu, X., Kha, L.-C. T., Zhu, X., Ryan, O., Jiang, Z., Zacksenhaus, E. (2005). Conserved and specific functions of mammalian ssu72. Nucleic Acids Res 33: 464-477 [Abstract] [Full Text]  
• Tsai, T., Fulton, L., Smith, B. J., Mueller, R. L., Gonzalez, G. A., Uusitalo, M. S., O'Brien, J. M. (2004). Rapid Identification of Germline Mutations in Retinoblastoma by Protein Truncation Testing. Arch Ophthalmol 122: 239-248 [Abstract] [Full Text]  
• Angus, S. P., Solomon, D. A., Kuschel, L., Hennigan, R. F., Knudsen, E. S. (2003). Retinoblastoma Tumor Suppressor: Analyses of Dynamic Behavior in Living Cells Reveal Multiple Modes of Regulation. Mol. Cell. Biol. 23: 8172-8188 [Abstract] [Full Text]  
• Markiewicz, E., Dechat, T., Foisner, R., Quinlan, Roy. A, Hutchison, C. J. (2002). Lamin A/C Binding Protein LAP2alpha Is Required for Nuclear Anchorage of Retinoblastoma Protein. Mol. Biol. Cell 13: 4401-4413 [Abstract] [Full Text]  
• Yang, H., Williams, B. O., Hinds, P. W., Shih, T. S., Jacks, T., Bronson, R. T., Livingston, D. M. (2002). Tumor Suppression by a Severely Truncated Species of Retinoblastoma Protein. Mol. Cell. Biol. 22: 3103-3110 [Abstract] [Full Text]  
• Kishima, Y., Yamamoto, H., Izumoto, Y., Yoshida, K., Enomoto, H., Yamamoto, M., Kuroda, T., Ito, H., Yoshizaki, K., Nakamura, H. (2002). Hepatoma-derived Growth Factor Stimulates Cell Growth after Translocation to the Nucleus by Nuclear Localization Signals. J. Biol. Chem. 277: 10315-10322 [Abstract] [Full Text]  
• Chestukhin, A., Litovchick, L., Rudich, K., DeCaprio, J. A. (2002). Nucleocytoplasmic Shuttling of p130/RBL2: Novel Regulatory Mechanism. Mol. Cell. Biol. 22: 453-468 [Abstract] [Full Text]  
• Harbour, J. W. (2001). Molecular Basis of Low-Penetrance Retinoblastoma. Arch Ophthalmol 119: 1699-1704 [Abstract] [Full Text]  
• Paggi, M. G., Giordano, A. (2001). Who Is the Boss in the Retinoblastoma Family? The Point of View of Rb2/p130, the Little Brother. Cancer Res. 61: 4651-4654 [Abstract] [Full Text]  
• Cinti, C., Claudio, P. P., Howard, C. M., Neri, L. M., Fu, Y., Leoncini, L., Tosi, G. M., Maraldi, N. M., Giordano, A. (2000). Genetic Alterations Disrupting the Nuclear Localization of the Retinoblastoma-related Gene RB2/p130 in Human Tumor Cell Lines and Primary Tumors. Cancer Res. 60: 383-389 [Abstract] [Full Text]  
• Liang, S.-H., Clarke, M. F. (1999). A Bipartite Nuclear Localization Signal Is Required for p53 Nuclear Import Regulated by a Carboxyl-terminal Domain. J. Biol. Chem. 274: 32699-32703 [Abstract] [Full Text]  
• Gascard, P., Nunomura, W., Lee, G., Walensky, L. D., Krauss, S. W., Takakuwa, Y., Chasis, J. A., Mohandas, N., Conboy, J. G. (1999). Deciphering the Nuclear Import Pathway for the Cytoskeletal Red Cell Protein 4.1R. Mol. Biol. Cell 10: 1783-1798 [Abstract] [Full Text]  
• Efthymiadis, A., Shao, H., Hubner, S., Jans, D. A. (1997). Kinetic Characterization of the Human Retinoblastoma Protein Bipartite Nuclear Localization Sequence (NLS) in Vivo and in Vitro. A COMPARISON WITH THE SV40 LARGE T-ANTIGEN NLS. J. Biol. Chem. 272: 22134-22139 [Abstract] [Full Text]  
• Zacksenhaus, E, Jiang, Z, Chung, D, Marth, J D, Phillips, R A, Gallie, B L (1996). pRb controls proliferation, differentiation, and death of skeletal muscle cells and other lineages during embryogenesis.. Genes Dev. 10: 3051-3064 [Abstract]  
• Sock, E., Enderich, J., Rosenfeld, M. G., Wegner, M. (1996). Identification of the Nuclear Localization Signal of the POU Domain Protein Tst-1/Oct6. J. Biol. Chem. 271: 17512-17518 [Abstract] [Full Text]  
• Jans, D. A., Moll, T., Nasmyth, K., Jans, P. (1995). Cyclin-dependent Kinase Site-regulated Signal-dependent Nuclear Localization of the SWI5 Yeast Transcription Factor in Mammalian Cells. J. Biol. Chem. 270: 17064-17067 [Abstract] [Full Text]  
• Wang, Y., MacDonald, J. I. S., Kent, C. (1995). Identification of the Nuclear Localization Signal of Rat Liver CTP:Phosphocholine Cytidylyltransferase. J. Biol. Chem. 270: 354-360 [Abstract] [Full Text]  
• Welch, P J, Wang, J Y (1995). Disruption of retinoblastoma protein function by coexpression of its C pocket fragment.. Genes Dev. 9: 31-46 [Abstract]  
• Panda, D. K., Miao, D., Lefebvre, V., Hendy, G. N., Goltzman, D. (2001). The Transcription Factor SOX9 Regulates Cell Cycle and Differentiation Genes in Chondrocytic CFK2 Cells. J. Biol. Chem. 276: 41229-41236 [Abstract] [Full Text]  
• Jiang, Z., Zacksenhaus, E. (2002). Activation of retinoblastoma protein in mammary gland leads to ductal growth suppression, precocious differentiation, and adenocarcinoma. J. Cell Biol. 156: 185-198 [Abstract] [Full Text]