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Normal and Expanded Huntington’s Disease Gene Alleles Produce Distinguishable Proteins Due to Translation Across the CAG Repeat

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Abstract

Background

An expanded CAG trinucleotide repeat is the genetic trigger of neuronal degeneration in Huntington’s disease (HD), but its mode of action has yet to be discovered. The sequence of the HD gene places the CAG repeat near the 5′ end in a region where it may be translated as a variable polyglutamine segment in the protein product, huntingtin.

Materials and Methods

Antisera directed at amino acid stretches predicted by the DNA sequence upstream and downstream of the CAG repeat were used in Western blot and immunohistochemical analyses to examine huntingtin expression from the normal and the HD allele in lymphoblastoid cells and postmortem brain tissue.

Results

CAG repeat segments of both normal and expanded HD alleles are indeed translated, as part of a discrete ∼350-kD protein that is found primarily in the cytosol. The difference in the length of the N-terminal polyglutamine segment is sufficient to distinguish normal and HD huntingtin in a Western blot assay.

Conclusions

The HD mutation does not eliminate expression of the HD gene but instead produces an altered protein with an expanded polyglutamine stretch near the N terminus. Thus, HD pathogenesis is probably triggered by an effect at the level of huntingtin protein.

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Acknowledgments

We thank Drs. J. Settleman, S. Pillai, V. Ramesh, E. Harlow, A. Bernards, S. Reeves, and H. T. M. Timmers for supplying control antisera and for helpful discussion and the Brain Tissue Resource Center (McLean Hospital) for postmortem tissues and clinical information. This work was supported by National Institutes of Health Grants NS16367, NS22031, and NS32765, and by grants from Bristol-Myers Squibb, Inc., the Hereditary Disease Foundation, and the Huntington’s Disease Society of America. CA was supported by the Andrew B. Cogan Fellowship of the Hereditary Disease Foundation, and SM and MD were supported by fellowships from the Huntington’s Disease Society of America and the Hereditary Disease Foundation, respectively.

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Authors and Affiliations

  1. Molecular Neurogenetics Unit, Massachusetts General Hospital East, Building 149, 13th Street, Charlestown, Massachusetts, 02129-2060, USA

    Francesca Persichetti, Christine M. Ambrose, Sandra M. McNeil, Jayalakshmi Srinidhi, Mary Anne Anderson, Barbara Jenkins, Glenn T. Barnes, Mabel P. Duyao, Marcy E. MacDonald & James F. Gusella

  2. Laboratory for Molecular Neuropathology, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    Pei Ge & Jean-Paul Vonsattel

  3. Brain Tissue Resource Center, McLean Hospital, Belmont, Massachusetts, USA

    Lisa Kanaley & Edward D. Bird

  4. Hereditary Disease Foundation, Santa Monica, California, USA

    Nancy S. Wexler

  5. Department of Neurology, Boston University Medical School, Boston, Massachusetts, USA

    Richard H. Myers

  6. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    James F. Gusella

Authors
  1. Francesca Persichetti
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  2. Christine M. Ambrose
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  3. Pei Ge
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  4. Sandra M. McNeil
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  5. Jayalakshmi Srinidhi
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  6. Mary Anne Anderson
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  7. Barbara Jenkins
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  8. Glenn T. Barnes
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  9. Mabel P. Duyao
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  10. Lisa Kanaley
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  11. Nancy S. Wexler
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  12. Richard H. Myers
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  13. Edward D. Bird
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  14. Jean-Paul Vonsattel
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  15. Marcy E. MacDonald
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  16. James F. Gusella
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  17. S. H. Orkin
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Persichetti, F., Ambrose, C.M., Ge, P. et al. Normal and Expanded Huntington’s Disease Gene Alleles Produce Distinguishable Proteins Due to Translation Across the CAG Repeat. Mol Med 1, 374–383 (1995). https://doi.org/10.1007/BF03401575

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  • DOI: https://doi.org/10.1007/BF03401575

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