Affiliations: [a] Institute of Medical Genetics, Cardiff University, Heath Park, Cardiff, UK | [b] Institute of Psychological Medicine and Clinical Neuroscience, MRC Centre for Neuropsychiatric Genetics and Genomics, Hadyn Ellis Building, Cardiff University, UK | [c] Cardiff School of Biosciences, Sir Martin Evans Building, Museum Avenue, Cardiff, UK
Correspondence to: Alis C. Hughes, Institute of Psychological Medicine and Clinical Neuroscience, MRC Centre for Neuropsychiatric Genetics and Genomics, Hadyn Ellis Building, Cardiff University, CF24 4HQ. Tel.: +44 2920 688 472; Fax: +44 2920 687 068; [email protected]
Note:  These authors contributed equally to this work.
Background: A CAG repeat expansion in HTT has been known to cause Huntington’s disease for over 20 years. The genomic sequence of the 67 exon HTT is clear but few reports have detailed alternative splicing or alternative transcripts. Most eukaryotic genes with multiple exons show alternative splicing that increases the diversity of the transcriptome and proteome: it would be surprising if a gene with 67 known exons in its two major transcripts did not present some alternative transcripts.
Objective: To investigate the presence of alternative transcripts directly in human HTT.
Methods: An overlapping RT-PCR based approach was used to determine novel HTT splice variants in human brain from HD patients and controls and 3D protein homology modelling employed to investigate their significance on the function of the HTT protein.
Results: Here we show multiple previously unreported novel transcripts of HTT. Of the 22 splice variants found, eight were in-frame with the potential to encode novel HTT protein isoforms. Two splice variants were selected for further study; HTT
Δex4,5,6 which results in the skipping of exons 4, 5 and 6 and HTTex41b which includes a novel exon created via partial retention of intron 41. 3D protein homology modelling showed that both splice variants are of potential functional significance leading to the loss of a karyopherin nuclear localisation signal and alterations to sites of posttranslational modification.
Conclusions: The identification of novel HTT transcripts has implications for HTT protein isoform expression and function. Understanding the functional significance of HTT alternative splicing would be critical to guide the design of potential therapeutics in HD that aim to reduce the toxic HTT transcript or protein product including RNA silencing and correction of mis-splicing in disease.
Keywords: Huntington’s disease, alternative splicing, RNA species, protein isoforms, structural modelling