Genomic organization of the rat pituitary adenylate cyclase-activating polypeptide receptor gene. Alternative splicing within the 5'-untranslated region

Tapan Kumar Chatterjee, Xuebo Liu, Robin L. Davisson, Rory A. Fisher

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP) elicits its diverse biological actions by interacting with both PACAP-selective type I PACAP receptors (PACAPRs) and type II PACAPRs that do not distinguish between PACAP and vasoactive intestinal polypeptide. Using long distance polymerase chain reaction, we amplified and characterized the entire coding region of the rat type I PACAPR (rPACAPR) gene, which spans 40 kilobases and contains 15 exons. Mapping of the exons and sequencing of all intron-exon boundaries revealed a structural organization of the rPACAPR gene that is very similar to those encoding other members of the calcitonin/secretin/parathyroid hormone receptor family. Southern blot analysis demonstrated a single copy of the rPACAPR gene. A combination of rapid amplification of cDNA ends and reverse transcriptase polymerase chain reaction revealed an unexpected diversity in the rPACAPR mRNA in the 5'-untranslated (5'-UTR) region. Four rPACAPR cDNAs were identified with 5'-UTR sequences that all diverged from the genomic sequence at a site 76 bp upstream of the ATG start codon, where a consensus 8' slice acceptor sequence was located. Sequence analysis of these amplified transcripts demonstrated that they arise by tissue-specific differential usage of four exons in the 5' non-coding region of the rPACAPR gene. This study is the first to elucidate the structural organization of a PACAPR gene and to demonstrate that alternative splicing generates rPACAPR transcripts with unique 5'-UTRs.

Original languageEnglish (US)
Pages (from-to)12122-12131
Number of pages10
JournalJournal of Biological Chemistry
Volume272
Issue number18
DOIs
StatePublished - May 2 1997

Fingerprint

Pituitary Adenylate Cyclase-Activating Polypeptide Receptors
5' Untranslated Regions
Alternative Splicing
Rats
Genes
Pituitary Adenylate Cyclase-Activating Polypeptide
Exons
Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Polymerase chain reaction
Receptors, Vasoactive Intestinal Polypeptide, Type I
Complementary DNA
Parathyroid Hormone Receptor Type 1
Secretin
Initiator Codon
Vasoactive Intestinal Peptide
RNA-Directed DNA Polymerase
Calcitonin
Southern Blotting
Reverse Transcriptase Polymerase Chain Reaction
Introns

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Genomic organization of the rat pituitary adenylate cyclase-activating polypeptide receptor gene. Alternative splicing within the 5'-untranslated region. / Chatterjee, Tapan Kumar; Liu, Xuebo; Davisson, Robin L.; Fisher, Rory A.

In: Journal of Biological Chemistry, Vol. 272, No. 18, 02.05.1997, p. 12122-12131.

Research output: Contribution to journalArticle

Chatterjee, Tapan Kumar ; Liu, Xuebo ; Davisson, Robin L. ; Fisher, Rory A. / Genomic organization of the rat pituitary adenylate cyclase-activating polypeptide receptor gene. Alternative splicing within the 5'-untranslated region. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 18. pp. 12122-12131.
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