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The nuclear location signal.

A E Smith, D Kalderon, B L Roberts

    Proceedings of the Royal Society of London. Series B, Biological Sciences
    |October 22, 1985
    PubMed
    Summary
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    A specific amino acid sequence in SV40 Large T protein acts as a nuclear localization signal, directing proteins to the nucleus. This signal

    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Virology

    Background:

    • Nuclear localization signals (NLS) are crucial for directing proteins to the cell nucleus.
    • The Simian virus 40 (SV40) Large T antigen is a well-studied viral oncoprotein with a known nuclear function.
    • Understanding NLS sequences is fundamental to comprehending protein trafficking and cellular regulation.

    Purpose of the Study:

    • To identify and characterize the specific amino acid sequence responsible for the nuclear localization of SV40 Large T antigen.
    • To determine if this sequence is sufficient to confer nuclear localization upon other proteins.
    • To investigate the presence and potential function of similar sequences in other proteins.

    Main Methods:

    • Site-directed mutagenesis was used to alter specific amino acids within the SV40 Large T sequence.

    Related Experiment Videos

  • Subcellular localization of wild-type and mutant proteins was assessed.
  • The identified sequence was experimentally transposed into heterologous proteins (Escherichia coli beta-galactosidase, chicken muscle pyruvate kinase) to test its function.
  • Bioinformatic analysis was performed to search for similar sequences in a large protein database.

    • Main Results:

    • A short sequence (Pro-Pro-Lys-Lys-Lys-Arg-Lys-Val) rich in basic amino acids within SV40 Large T was identified as essential for nuclear localization.
    • Mutations within this sequence resulted in cytoplasmic localization of the Large T protein.
    • Transposition of this sequence to the N-terminus of cytoplasmic proteins successfully directed them to the nucleus, confirming its function as a potent NLS.
    • Similar basic sequences were found in other nuclear proteins, but also in non-nuclear proteins, suggesting context-dependent functionality.

    Conclusions:

    • The identified basic amino acid-rich sequence functions as a sufficient nuclear localization signal (NLS).
    • This NLS can direct heterologous proteins to the nucleus, highlighting its versatility.
    • The presence and accessibility of similar sequences in other proteins may determine their nuclear or cytoplasmic localization, with potential implications for protein function.