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An Escherichia coli-Based Genetic Strategy for Characterizing RNA Binding Proteins

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RNA-Protein Interaction Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 118))

Abstract

In recent years, interest in RNA-binding proteins (RNA-BPs) and RNA-protein interactions has grown considerably. An important reason for this interest is the increased recognition that RNA-BPs are involved in a wide variety of critical viral and cellular processes including transcription, translation, RNA processing, RNA transport, gene regulation, and splicing. A second reason lies in the technical advances and the generation of new methods that have simplified the analysis of RNA-protein interactions. Nevertheless, a thorough analysis of an RNA-BP is not a trivial task, and, at present, there are only a few RNA-BPs that have been characterized in detail.

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References

  1. Jain, C. and Belasco, J. G. (1996) A structural model for the HIV-1 Rev-RRE complex deduced from the altered specificity of Rev variants isolated by a rapid genetic screening strategy. Cell 87, 115–125.

    Article  PubMed  CAS  Google Scholar 

  2. Bouvet, P., Jain, C., Belasco, J. G., Almaric, F., and Erard, M. (1997) RNA recognition by the joint action of two nucleolin RNA-binding domains: genetic analysis and structural modeling. EMBO J. 16, 5235–5246.

    Article  PubMed  CAS  Google Scholar 

  3. Dreyfus, M. (1988) What constitutes the signal for the initiation of protein synthesis on E. coli mRNAs? J. Mol. Biol. 204, 79–94.

    Article  PubMed  CAS  Google Scholar 

  4. Gold, L. (1988) Posttranscriptional regulatory mechanisms in Escherichia coli. Annu. Rev. Biochem. 57, 199–233.

    Article  PubMed  CAS  Google Scholar 

  5. de Smit, M. H. and van Duin, J. (1990) Secondary structure of the ribosome binding site determines translational efficiency: a quantitative analysis. Proc. Natl. Acad. Sci. USA 87, 7668–7672.

    Article  PubMed  Google Scholar 

  6. Rose, R. E. (1988) The nucleotide sequence of pACYC184. Nucleic Acids. Res. 16, 355.

    Article  PubMed  CAS  Google Scholar 

  7. Miller, J. H. (1972) in Experiments in Molecular Genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  8. Peabody, D. (1993) The RNA binding site of bacteriophage MS2 coat protein. EMBO J. 12, 595–600.

    PubMed  CAS  Google Scholar 

  9. Kunkel, T. A., Bebenek, K., and McClary, J. (1991) Efficient site-directed mutagenesis using uracil-containing DNA. Methods Enzymol. 204, 125–139.

    Article  PubMed  CAS  Google Scholar 

  10. Olsen, D. B., Sayers, J. R., and Eckstein, F. (1993) Site-directed mutagenesis of single-stranded and double-stranded DNA by phosphorothionate approach. Methods Enzymol. 217, 189–217.

    Article  PubMed  CAS  Google Scholar 

  11. Cadwell, R. C. and Joyce, G. F. (1992) Randomization of genes by PCR mutagenesis. PCR Methods Appl. 2, 28–33.

    PubMed  CAS  Google Scholar 

  12. Jain, C. and Belasco, J. G. (1997) A genetic method for the study of RNA-binding proteins, in mRNA Formation and Function (Richter, J., ed.), Academic Press, New York, NY, pp. 263–284.

    Chapter  Google Scholar 

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© 1999 Humana Press Inc.

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Jain, C. (1999). An Escherichia coli-Based Genetic Strategy for Characterizing RNA Binding Proteins. In: Haynes, S.R. (eds) RNA-Protein Interaction Protocols. Methods in Molecular Biology™, vol 118. Humana Press. https://doi.org/10.1385/1-59259-676-2:161

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  • DOI: https://doi.org/10.1385/1-59259-676-2:161

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-568-3

  • Online ISBN: 978-1-59259-676-8

  • eBook Packages: Springer Protocols

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