Characterisation of novel splicing variants of the tyrosine hydroxylase C-terminal domain in human neuroblastic tumours.
Alternative splicing of human tyrosine hydroxylase (hTH) transcripts appears to occur mainly in the N-terminal domain, giving rise to at least eight different isoforms. We recently reported the existence of hTH transcript variants resulting from splicing of exons 8 and 9, within a region previously thought to be constant. The mRNA distribution of these novel hTH isoforms in neuroblastic tumours and in foetal adrenal glands was analysed by conventional and real-time RT-PCR. The presence of the target protein was determined by Western blotting, immunoprecipitation and protein analysis. Transcripts lacking exons 8 and 9 were widely distributed in the tissues analysed. Characterisation of full-length mRNA revealed that splicing of exons 8 and 9 was always associated with splicing of exons 2 (hTH-Delta2,8,9) or 1b and 2 (hTH-Delta1b,2,8,9). In addition, one variant detected on Western blots in several tumours fits the predicted size (58 kDa) of the isoforms lacking exons 8 and 9. In conclusion, the two novel isoforms reported here (hTH-Delta2,8,9 and hTH-Delta1b,2,8,9) represent the first full-length isoforms with alternative splicing in the hTH C-terminal domain. These results demonstrate for the first time the existence of hTH isoforms Delta2,8,9 and Delta1b,2,8,9. Their general distribution in neuroblastoma and adrenal glands and translation into protein suggest a significant functional role for these novel hTH isoforms, which merit further study.
Roma
Josep
J
Biomedical Research Unit, Institut de Recerca, Hospital Universitari Vall d'Hebron, P Vall d'Hebron 119, E-08035 Barcelona, Spain.
Saus
Ester
E
Cuadros
Marc
M
Reventós
Jaume
J
Sánchez de Toledo
Josep
J
Gallego
Soledad
S
D000293:Adolescent; D017398:Alternative Splicing / Q000235:genetics*; D015153:Blotting, Western; D002648:Child; D002675:Child, Preschool; D005091:Exons / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007527:Isoenzymes / Q000235:genetics; D009447:Neuroblastoma / Q000201:enzymology*; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*
D007527:Isoenzymes; D011994:Recombinant Proteins; D014446:Tyrosine 3-Monooxygenase