Deregulation of alternative processing of Calcitonin/CGRP-I pre-mRNA by a single point mutation.
The Calcitonin/CGRP-I (CALC-I) gene was one of the first examples of a cellular gene exhibiting alternative, tissue-specific processing of its primary transcript. Calcitonin (CT) mRNA is the predominant product in thyroid C-cells, whereas CGRP-I (Calcitonin Gene Related Peptide-I) mRNA is the main product in neurons of the central and peripheral nervous systems. Investigating the molecular mechanism underlying the alternative processing events, we have demonstrated that the CT-specific splice acceptor site is an intrinsical weak site due to usage of a uridine branch acceptor. The data presented in this report show that a single point mutation changing the uridine branch acceptor into a commonly preferred adenosine residue results in the predominant production of CT mRNA in otherwise CGRP-I mRNA-producing F9 cells. The results of the experiments implicate that the low efficiency of CT splicing, caused by usage of a uridine branch acceptor, allows the production of CGRP-I mRNA in neural cells.
Adema
G J
GJ
Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, The Netherlands.
Baas
P D
PD
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002116:Calcitonin / Q000235:genetics*; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics*; D005091:Exons; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D013724:Teratoma; D014158:Transcription, Genetic; D014162:Transfection
D015345:Oligonucleotide Probes; D012322:RNA Precursors; D012333:RNA, Messenger; D002116:Calcitonin; D015740:Calcitonin Gene-Related Peptide