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Isolation and functional expression of pituitary peptidylglycine alpha-amidating enzyme mRNA. A variant lacking the

I Kato1, H Yonekura, H Yamamoto

  • 1Department of Biochemistry, Tohoku University School of Medicine, Miyagi, Japan.

FEBS Letters
|September 3, 1990
PubMed

Insights

Alternative splicing creates multiple messenger RNAs (mRNAs) for peptidylglycine alpha-amidating enzyme in rat pituitary. A secreted form is efficiently produced by a variant mRNA lacking a transmembrane domain.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Endocrinology

Background:

  • Peptidylglycine alpha-amidating enzyme (PAOE) is crucial for producing biologically active peptides.
  • Understanding PAOE gene expression is vital for peptide hormone regulation.

Purpose of the Study:

  • To investigate the diversity of mRNA transcripts encoding PAOE in the rat pituitary.
  • To identify the functional significance of different PAOE mRNA variants.

Main Methods:

  • Southern blot analysis to detect gene structure.
  • Ribonuclease protection assays to analyze mRNA species.
  • cDNA expression in COS-7 cells to assess enzyme function.

Main Results:

  • At least five distinct mRNAs encode PAOE in rat pituitary.
  • Alternative splicing is responsible for generating these mRNA variants.
  • A major pituitary mRNA variant lacks the transmembrane domain-coding sequence.
  • This variant efficiently produces a secreted 37 kDa form of the enzyme when expressed.

Conclusions:

  • Alternative splicing generates functionally distinct PAOE mRNA variants in the rat pituitary.
  • The transmembrane domain-lacking variant is a key transcript for producing secreted PAOE.
  • This finding has implications for understanding peptide hormone processing and secretion.

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