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Changes in globin messenger RNA content during erythroid cell differentiation.

F Ramirez, R Gambino, G M Maniatis

    The Journal of Biological Chemistry
    |August 10, 1975
    PubMed
    Summary

    Erythroid cell differentiation involves a significant increase in globin messenger RNA (mRNA) content, particularly after erythropoietin stimulation. This study quantifies globin mRNA accumulation during erythropoiesis.

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    Area of Science:

    • Hematology
    • Molecular Biology
    • Cellular Differentiation

    Background:

    • Erythroid precursor cells initially show minimal globin synthesis and globin mRNA.
    • Erythropoietin (EPO) stimulation leads to increased globin synthesis and translatable globin mRNA.
    • Direct measurement of globin mRNA sequences during differentiation was previously lacking.

    Purpose of the Study:

    • To directly quantify globin mRNA sequences during erythroid cell differentiation.
    • To investigate the kinetics of globin mRNA accumulation in response to erythropoietin.
    • To determine the presence of an untranslated globin mRNA pool in precursor cells.

    Main Methods:

    • Isolation of mouse fetal erythroid precursor cells.
    • Culture of cells with erythropoietin.

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  • Molecular hybridization using 3H-labeled complementary DNA (cDNA) to globin mRNA.
  • Measurement of globin mRNA content via cDNA:mRNA hybridization kinetics (Cot1/2 values).

    • Main Results:

    • Erythroid precursor cells contain very few globin mRNA sequences.
    • No significant pool of untranslated globin mRNA was detected.
    • Globin mRNA content increased 250-fold within 22 hours of EPO culture, with a further 2-fold increase by 44 hours.
    • Cell numbers increased 2-3 fold during 44 hours of culture, reaching ~1800 globin mRNA molecules/cell by 22 hours.

    Conclusions:

    • Erythroid differentiation involves a substantial, time-dependent increase in globin mRNA levels.
    • Erythropoietin is crucial for inducing globin mRNA accumulation and subsequent globin synthesis.
    • Untranslated globin mRNA is not stored in significant amounts in early erythroid precursors.