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Complementation between LLC-PK1 mutants affected in polypeptide hormone-receptor function.

D A Jans, T J Resink, B A Hemmings

    European Journal of Biochemistry
    |February 2, 1987
    PubMed
    Summary
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    Mutant cell lines showed reduced calcitonin receptor function but increased vasopressin receptor numbers. These defects are recessive and affect the same gene, suggesting a common pathway for polypeptide hormone receptor processing.

    Area of Science:

    • Cell Biology
    • Endocrinology
    • Molecular Genetics

    Background:

    • Mutant LLC-PK1 cell lines (FIB6, FIB5/N4) were studied to understand polypeptide hormone receptor regulation.
    • Previous studies indicated altered responses to calcitonin and vasopressin in these mutants.

    Purpose of the Study:

    • To investigate the specific defects in calcitonin and vasopressin receptor function in FIB6 and FIB5/N4 cell lines.
    • To determine the genetic basis and inheritance pattern of these receptor defects.
    • To explore potential common pathways in the processing of calcitonin and vasopressin receptors.

    Main Methods:

    • Hormone binding assays using radiolabeled calcitonin and vasopressin.
    • Measurement of adenylate cyclase and cAMP-dependent protein kinase (cAMP-PK) activation.

    Related Experiment Videos

  • Somatic cell hybridization to assess complementation of mutant phenotypes.
  • Analysis of receptor expression and function in parental, mutant, and hybrid cell lines.

    • Main Results:

    • Mutant cell lines exhibited significantly reduced calcitonin binding and activation of downstream signaling pathways.
    • Concurrently, mutants showed increased vasopressin binding and receptor responsiveness.
    • Somatic cell hybrids with parental cells restored normal receptor function, indicating recessive mutations.
    • Hybridization between FIB6 and FIB5/N4 showed no complementation, suggesting mutation in the same gene.
    • Hybridization with a 'receptorless' mutant (M18) demonstrated complementation, indicating distinct defects.

    Conclusions:

    • The mutations in FIB6 and FIB5/N4 are recessive and affect the same gene involved in polypeptide hormone receptor function.
    • Mutant cell lines possess an increased number of functional vasopressin receptors, possibly due to compensatory mechanisms.
    • The findings support a model proposing a common pathway for the processing and regulation of calcitonin and vasopressin receptors.