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Cooperation of 5' and 3' processing sites as well as intron and exon sequences in calcitonin exon recognition.
Nucleic Acids Res. 1995 Jan 25; 23(2):248-55.NA

Abstract

We have previously shown that the calcitonin (CT)-encoding exon 4 of the human calcitonin/calcitonin gene-related peptide I (CGRP-I) gene (CALC-I gene) is surrounded by suboptimal processing sites. At the 5' end of exon 4 a weak 3' splice site is present because of an unusual branch acceptor nucleotide (U) and a weak poly(A) site is present at the 3' end of exon 4. For CT-specific RNA processing two different exon enhancer elements, A and B, located within exon 4 are required. In this study we have investigated the cooperation of these elements in CT exon recognition and inclusion by transient transfection into 293 cells of CALC-I minigene constructs. Improvement of the strength of the 3' splice site in front of exon 4 by the branchpoint mutation U-->A reduces the requirement for the presence of exon enhancer elements within exon 4 for CT-specific RNA processing, irrespective of the length of exon 4. Replacement of the exon 4 poly(A) site with a 5' splice site does not result in CT exon recognition, unless also one or more exon enhancer elements and/or the branchpoint mutation U-->A in front of exon 4 are present. This indicates that terminal and internal exons are recognised in a similar fashion. The number of additional enhancing elements that are required for CT exon recognition depends on the strength of the 5' splice site. Deletion of a large part of intron 4 also leads to partial exon 4 skipping. All these different elements contribute to CT exon recognition and inclusion. The CT exon is recognised as a whole entity and the sum of the strengths of the different elements determines recognition as an exon. Curiously, in one of our constructs a 5' splice site at the end of exon 4 is either ignored by the splicing machinery of the cell or recognised as a splice donor or as a splice acceptor site.

Authors+Show Affiliations

Laboratory for Physiological Chemistry, Utrecht University, Netherlands.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

7862529

Citation

Zandberg, H, et al. "Cooperation of 5' and 3' Processing Sites as Well as Intron and Exon Sequences in Calcitonin Exon Recognition." Nucleic Acids Research, vol. 23, no. 2, 1995, pp. 248-55.
Zandberg H, Moen TC, Baas PD. Cooperation of 5' and 3' processing sites as well as intron and exon sequences in calcitonin exon recognition. Nucleic Acids Res. 1995;23(2):248-55.
Zandberg, H., Moen, T. C., & Baas, P. D. (1995). Cooperation of 5' and 3' processing sites as well as intron and exon sequences in calcitonin exon recognition. Nucleic Acids Research, 23(2), 248-55.
Zandberg H, Moen TC, Baas PD. Cooperation of 5' and 3' Processing Sites as Well as Intron and Exon Sequences in Calcitonin Exon Recognition. Nucleic Acids Res. 1995 Jan 25;23(2):248-55. PubMed PMID: 7862529.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cooperation of 5' and 3' processing sites as well as intron and exon sequences in calcitonin exon recognition. AU - Zandberg,H, AU - Moen,T C, AU - Baas,P D, PY - 1995/1/25/pubmed PY - 1995/1/25/medline PY - 1995/1/25/entrez SP - 248 EP - 55 JF - Nucleic acids research JO - Nucleic Acids Res VL - 23 IS - 2 N2 - We have previously shown that the calcitonin (CT)-encoding exon 4 of the human calcitonin/calcitonin gene-related peptide I (CGRP-I) gene (CALC-I gene) is surrounded by suboptimal processing sites. At the 5' end of exon 4 a weak 3' splice site is present because of an unusual branch acceptor nucleotide (U) and a weak poly(A) site is present at the 3' end of exon 4. For CT-specific RNA processing two different exon enhancer elements, A and B, located within exon 4 are required. In this study we have investigated the cooperation of these elements in CT exon recognition and inclusion by transient transfection into 293 cells of CALC-I minigene constructs. Improvement of the strength of the 3' splice site in front of exon 4 by the branchpoint mutation U-->A reduces the requirement for the presence of exon enhancer elements within exon 4 for CT-specific RNA processing, irrespective of the length of exon 4. Replacement of the exon 4 poly(A) site with a 5' splice site does not result in CT exon recognition, unless also one or more exon enhancer elements and/or the branchpoint mutation U-->A in front of exon 4 are present. This indicates that terminal and internal exons are recognised in a similar fashion. The number of additional enhancing elements that are required for CT exon recognition depends on the strength of the 5' splice site. Deletion of a large part of intron 4 also leads to partial exon 4 skipping. All these different elements contribute to CT exon recognition and inclusion. The CT exon is recognised as a whole entity and the sum of the strengths of the different elements determines recognition as an exon. Curiously, in one of our constructs a 5' splice site at the end of exon 4 is either ignored by the splicing machinery of the cell or recognised as a splice donor or as a splice acceptor site. SN - 0305-1048 UR - https://www.unboundmedicine.com/medline/citation/7862529/Cooperation_of_5'_and_3'_processing_sites_as_well_as_intron_and_exon_sequences_in_calcitonin_exon_recognition_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/7862529/ DB - PRIME DP - Unbound Medicine ER -