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Identification and cloning of the cDNA of a Rb-associated protein RAP140a

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Abstract

Rb exerts important physiological functions in cell-cycle control, gene expression, cell differentiation, apoptosis, development and tumorigenesis by interacting with many cellular proteins. Using human partial Rb as bait, we screened a human fetal brain cDNA library through yeast two-hybrid system and obtained six novel cDNA fragments. Among them, one cDNA fragment corresponds to two different transcripts, 7 kb and 9 kb in Northern blot analysis. These two transcripts showed uniform distribution in various human tissues. We cloned the full-length cDNA of a 7.2 kb transcript through three times PCR amplifications. It was namedRAP140a and predicted to encode a 1 233 amino acids hydrophilic protein.RAP140a was mapped to chromosome 3p13-p14. 1. RAP140a may be functionally related to the intracellular translocation of Rb or other proteins.

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References

  1. Weinberg, R. A., The retinoblastoma protein and cell cycle control, Cell, 1995, 81: 323.

    Article  PubMed  CAS  Google Scholar 

  2. Bartek, J., Bartkova, J., Lukas, J., The retinoblastoma protein pathway and the restriction point, Curr. Opin. Cell. Biol., 1996, 8: 805.

    Article  PubMed  CAS  Google Scholar 

  3. Mittnacht, S., Control of Rb phosphorylation, Curr. Opin. Genet. Devel., 1998, 8: 21.

    Article  CAS  Google Scholar 

  4. Macleod, K., Rb and E2F-1 in mouse development and tumorigenesis, Curr. Opin. Genet. Dev., 1999, 9: 31.

    Article  PubMed  CAS  Google Scholar 

  5. Chen, P. L., Riley, D. J., Chen, Y. et al., Retinoblastoma protein positively regulates terminal adipocyte differentiation through direct interactions with C/EBPs, Genes Dev., 1996, 10: 2794.

    Article  PubMed  CAS  Google Scholar 

  6. Luo, R. X., Postigo, A. A., Dean, D. C., Rb interacts with histone deacetylase to repress transcription, Cell, 1998, 92: 463.

    Article  PubMed  CAS  Google Scholar 

  7. Fields, S., Song, O., A novel genetic system to detect protein-protein interactions, Nature, 1989, 340: 245.

    Article  PubMed  CAS  Google Scholar 

  8. Fields, S., Sternglanz, R., The two-hybrid system: an assay for protein-protein interactions, Trends Genet., 1994, 10: 286.

    Article  PubMed  CAS  Google Scholar 

  9. Feilotter, H. E., Hannon, G. J., Ruddel, C. J. et al., Construction of an improved host strain for two hybrid screening, Nucleic Acid Res., 1994, 22: 1502.

    Article  PubMed  CAS  Google Scholar 

  10. Van Aelst, L., Barr, M., Marcus, S. et al., Complex formation between RAS and RAF and other protein kinases, Proc. Natl. Acad. Sci. USA, 1993, 90: 6213.

    Article  PubMed  Google Scholar 

  11. Bartel, P. L., Chien, C. T., Sternglanz, R., Elimination of false positives that arise in using the two-hybrid system, Bio-Techniques, 1993, 14: 920.

    CAS  Google Scholar 

  12. Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.

    Google Scholar 

  13. He, K. L., Gu, B. W., Zhang, Q. H. et al., Application of radiation hybrid techniques in chromosomal localizations, Science in China (in Chinese), Ser. C, 1999, 29(1): 108.

    Google Scholar 

  14. Altschul, S. F., Gish, W., Miller, W. et al., Basic local alignment search tool, J. Mol. Biol., 1990, 215(3): 403.

    PubMed  CAS  Google Scholar 

  15. Kyte, J., Doolittle, R. F., A method for displaying the hydrophobic character of a protein, J. Mol. Biol., 1982, 157: 105.

    Article  PubMed  CAS  Google Scholar 

  16. Chou, P. Y., Fasman, G. D., Prediction of the secondary structure of proteins from their amino acid sequence, Adv. Enzyme Regul., 1978, 47:45.

    CAS  Google Scholar 

  17. Reinhardt, A., Hubbard, T., Using neural networks for prediction of the subcellular location of proteins, Nucleic Acids Res., 1998, 26(9): 2230.

    Article  PubMed  CAS  Google Scholar 

  18. Wang, J. Y. J., Knudsen, E. S., Welch, P. J., The retinoblastoma tumor suppress protein, Adv. Cancer Res., 1994, 64: 25.

    Article  PubMed  CAS  Google Scholar 

  19. Wang, J. Y. J., Retinoblastoma protein in growth suppression and death protection, Curr. Opin. Genet. Dev., 1997, 7: 39.

    Article  PubMed  CAS  Google Scholar 

  20. Fush, E., The cytoskeleton and disease: Genetic diseases of intermediate filament, Ann. Rev. of Genet., 1996, 30: 197.

    Article  Google Scholar 

  21. McLachlan, A. D., Karn, J., Periodic features in the amino acid sequence of nematode myosin rod, J. Mol. Biol., 1983, 164: 605.

    Article  PubMed  CAS  Google Scholar 

  22. Nakajima, H., Hirata, A., Ohawa, Y. et al., A cytoskeleton-related gene, USO1, is required for intracellular protein transport inSaccharomyces cerevisiae, J. Cell Biol., 1991, 113(2): 245.

    Article  PubMed  CAS  Google Scholar 

  23. Dornert, C., Ciossek, T., Muller, S. et al., Characterization of KIF1C, a new kinesin-like protein involved in vesicle transport from the Golgi apparatus to the endoplasmic reticulum, J. Biol. Chem., 1998, 273(32): 20267.

    Article  Google Scholar 

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Correspondence to Shizhou Ao.

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Li, Q., Wen, H. & Ao, S. Identification and cloning of the cDNA of a Rb-associated protein RAP140a. Sci. China Ser. C.-Life Sci. 43, 637–647 (2000). https://doi.org/10.1007/BF02882285

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  • DOI: https://doi.org/10.1007/BF02882285

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