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Cloning and sequencing of the cDNA encoding the avian kidney cAMP-dependent protein kinase inhibitor protein

G S Marchetto1, H L Henry

  • 1Department of Biochemistry, University of California, Riverside 92521, USA.

Gene
|June 9, 1995
PubMed

Insights

Vitamin D deficiency affects protein kinase inhibitor (PKI) levels in chick kidneys. Researchers isolated a chick PKI cDNA, revealing conserved inhibitory regions and unique C-terminal sequences compared to mammals.

Area of Science:

  • Molecular Biology
  • Endocrinology
  • Biochemistry

Background:

  • The endogenous inhibitor of cAMP-dependent protein kinase (PKA) is regulated by vitamin D status.
  • Vitamin D-replete chicks exhibit lower levels of PKA inhibitor in kidney tissue compared to vitamin D-deficient chicks.

Purpose of the Study:

  • To isolate and characterize the cDNA encoding the protein kinase inhibitor (PKI) from vitamin D-deficient chick kidneys.
  • To compare the deduced amino acid sequence of avian PKI with mammalian isoforms.

Main Methods:

  • Screening of a vitamin D-deficient chick kidney cDNA library.
  • Isolation and sequencing of a 450-bp cDNA clone encoding a 76-amino acid protein.
  • Bioinformatic analysis to compare amino acid sequences with mammalian PKI alpha and beta 1.

Main Results:

  • A cDNA clone encoding avian PKI was successfully isolated.
  • Avian PKI shares 80% and 41% amino acid identity with mammalian PKI alpha and PKI beta 1, respectively.
  • Conserved N-terminal sequences, including the pseudo-substrate site (18GRRNA22), are present in both avian and mammalian PKI.
  • Chick kidney PKI possesses ten unique amino acids in its C-terminal region not found in mammalian PKI alpha or beta.

Conclusions:

  • Vitamin D status influences the expression of endogenous PKA inhibitor in chick kidneys.
  • The identified avian PKI shares functional similarities with mammalian isoforms, particularly in the N-terminal inhibitory domain.
  • Unique C-terminal sequences in avian PKI may confer distinct regulatory or functional properties.

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