Unusual branch point selection involved in splicing of the alternatively processed Calcitonin/CGRP-I pre-mRNA.
To study splice site selection in alternative RNA processing we used the human Calcitonin/CGRP-I (CALC-I) gene. Expression of the CALC-I gene in thyroid C-cells results predominantly in calcitonin (CT) mRNA (containing exons 1 to 4) whereas CGRP-I mRNA (containing exons 1,2,3,5 and 6) is the exclusive product in particular nerve cells. We previously reported that a model precursor RNA containing the exon 3 to exon 5 region is predominantly processed into CGRP-I mRNA in vitro using nuclear extracts of three different cell types. To study CT specific processing in Hela cell nuclear extracts we have used precursor RNAs corresponding to the exon 3 to exon 4 region containing only CT specific processing signals. The results revealed the usage of a uridine residue 23 nucleotides upstream of the 3' splice site as the major site of lariat formation in CT specific splicing. The implications of this finding for the alternative, tissue specific processing of the CALC-I pre-mRNA and for branch point selection in general are discussed.
Adema
G J
GJ
Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, The Netherlands.
Bovenberg
R A
RA
Jansz
H S
HS
Baas
P D
PD
D002116:Calcitonin / Q000235:genetics*; D015740:Calcitonin Gene-Related Peptide; D005091:Exons; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D009479:Neuropeptides / Q000235:genetics*; D012315:RNA Caps / Q000378:metabolism; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*
D009479:Neuropeptides; D012315:RNA Caps; D012322:RNA Precursors; D012333:RNA, Messenger; D002116:Calcitonin; D015740:Calcitonin Gene-Related Peptide