Micro-processing events in mRNAs identified by DHPLC analysis

Angela Gallo, Emma Thomson, James Brindle, Mary A. O'Connell, Liam P. Keegan

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Post-transcriptional processes such as alternative splicing and RNA editing have a huge impact on the diversity of the proteome. Detecting alternatively spliced transcripts is difficult when they are rare. In addition, edited transcripts often differ from the genomic sequence by only a few nucleotides. Denaturing high performance liquid chromatography (DHPLC) is routinely used for single nucleotide polymorphism detection and we used this method to detect alternatively spliced or edited transcripts. As the sites of RNA editing appear to be conserved within gene families, we investigated whether editing sites are conserved in the murine homologue of the Drosophila cacophony transcript encoding the α1 subunit of a voltage-gated calcium channel that is edited at 10 independent positions. Although DHPLC analysis detects RNA editing at as low as 3% in control transcripts, no evidence of RNA editing was found in the analysed murine transcript. However an alternative exon was identified at the 3′ end of the mouse Cacna1α transcript and an alternative micro-exon encoding only two amino acids (NP) was found in the extracellular loop before the IVS4 helix in the same transcript. In the homologous Drosophila transcript a micro-exon also encoding two amino acids was found at the same position before the IVS4 helix.

Original languageEnglish
Pages (from-to)3945-3953
Number of pages9
JournalNucleic Acids Research
Volume30
Issue number18
Publication statusPublished - Sep 15 2002

Fingerprint

RNA Editing
High Pressure Liquid Chromatography
Exons
Messenger RNA
Drosophila
Amino Acids
Alternative Splicing
Proteome
Calcium Channels
Single Nucleotide Polymorphism
Nucleotides
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

Gallo, A., Thomson, E., Brindle, J., O'Connell, M. A., & Keegan, L. P. (2002). Micro-processing events in mRNAs identified by DHPLC analysis. Nucleic Acids Research, 30(18), 3945-3953.

Micro-processing events in mRNAs identified by DHPLC analysis. / Gallo, Angela; Thomson, Emma; Brindle, James; O'Connell, Mary A.; Keegan, Liam P.

In: Nucleic Acids Research, Vol. 30, No. 18, 15.09.2002, p. 3945-3953.

Research output: Contribution to journalArticle

Gallo, A, Thomson, E, Brindle, J, O'Connell, MA & Keegan, LP 2002, 'Micro-processing events in mRNAs identified by DHPLC analysis', Nucleic Acids Research, vol. 30, no. 18, pp. 3945-3953.
Gallo A, Thomson E, Brindle J, O'Connell MA, Keegan LP. Micro-processing events in mRNAs identified by DHPLC analysis. Nucleic Acids Research. 2002 Sep 15;30(18):3945-3953.
Gallo, Angela ; Thomson, Emma ; Brindle, James ; O'Connell, Mary A. ; Keegan, Liam P. / Micro-processing events in mRNAs identified by DHPLC analysis. In: Nucleic Acids Research. 2002 ; Vol. 30, No. 18. pp. 3945-3953.
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