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Decrease of renal phosphoenolpyruvate carboxykinase RNA and poly(A)+ RNA level by ochratoxin A.

H Meisner, M A Cimbala, R W Hanson

    Archives of Biochemistry and Biophysics
    |May 1, 1983
    PubMed
    Summary
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    Ochratoxin A reduces kidney phosphoenolpyruvate carboxykinase (PEPCK) mRNA levels in rats by decreasing total renal mRNA concentration, not by affecting transcription or translation efficiency.

    Area of Science:

    • Biochemistry
    • Toxicology
    • Molecular Biology

    Background:

    • Ochratoxin A (OA) is a nephrotoxin produced by Aspergillus ochraceus.
    • OA inhibits renal gluconeogenesis and decreases phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) activity in rat kidneys.

    Purpose of the Study:

    • To investigate the molecular mechanisms by which Ochratoxin A affects PEPCK expression in rat kidneys.
    • To determine if OA impacts mRNA levels, translation efficiency, or transcription rates.

    Main Methods:

    • Northern blotting and dot blot techniques were used to measure PEPCK mRNA sequence abundance.
    • Poly(A)+ RNA was isolated and translated in vitro to assess translational efficiency.
    • Nuclear transcription rates were measured using [32P]UTP incorporation.

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    Main Results:

    • Ochratoxin A significantly reduced translatable PEPCK mRNA levels in rat kidneys after 2 days of toxin feeding.
    • The efficiency of poly(A)+ RNA translation was not affected by OA.
    • Total poly(A)+ RNA levels in kidneys were reduced by 50% after 3-5 days of OA exposure, but hepatic levels remained unchanged.
    • Nuclear transcription rates of total RNA and PEPCK mRNA were not reduced by OA.
    • A 72,000 Mr protein mRNA species increased in relative abundance, but this protein was distinct from PEPCK.

    Conclusions:

    • Ochratoxin A lowers total renal mRNA concentration in rats.
    • PEPCK mRNA is reduced to a greater extent than the bulk of the renal RNA pool.
    • The mechanism involves decreased mRNA stability or processing, not altered transcription or translation efficiency.