Thyroid hormone decreases the stability and the poly(A) tract length of rat thyrotropin β-subunit messenger RNA

Ian M. Krane, Eliot Spindel, William W. Chin

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    Abstract

    Thyroid hormone (T3 and T4) down-regulation of TSH subunit steady state mRNA levels and subunit gene transcription in vitro and in vivo has been well studied. We present evidence here that T3 can also regulate the turnover of TSH subunit mRNA. The apparent half-life of the TSH β-subunit mRNA was determined by adding actinomycin-D (2 μM) to dispersed rat pituitary cultures in hypothyroid medium or medium containing 10-7 M T3 and analyzing the decline in subunit mRNA levels with time. The half-life of the TSHβ mRNA from those cultures treated with T3 was shorter than that of the control cultures (9 vs. >24 h, respectively). A possible mechanism by which TSH β-subunit mRNA stability is altered is through a change in the size of each mRNA's poly(A) tail. Northern blot analysis of total RNA from the above cultures revealed that T3 treatment reduces the size of the TSH β-subunit mRNA. To determine if this alteration of mRNA size was due to a loss of a portion of the poly(A) tract and not to alternative splicing of the transcript or use of a secondary transcriptional start site, pooled RNAs were hybridized with oligo(dT) and subsequently digested with RNAse-H to remove the poly(A) tract. RNA blot analysis of these RNAs showed that T3 treatment results in the loss of most of the TSHβ poly(A) tail. After 8 h in culture, the T3-induced shortening of the poly(A) tail is specific to the TSH β-subunit, as we saw no such effect on the similarly sized mRNAs (α-subunit, LHβ, or cyclophilin-A). The effect of T3 to shorten the TSHβ mRNA can be mimicked by cycloheximide (20 μg/ml) or actinomycin-D, indicating that a rapidly turning-over protein may be involved in maintaining the size and possibly the stability of the TSHβ mRNA under euthyroid and hypothyroid conditions. We have also observed that this T3-induced shortening of the TSHβ mRNA can be produced in vivo within 4 h after a single dose of T3 has been administered to euthyroid animals (20 μg/100 g BW, ip). Thus, T3 reduces the apparent half-life of the TSHβ mRNA and causes a concomitant decrease in its poly(A) tail size. Although a causal relationship has not been established, it is possible that shortening of the poly(A) tract may lead to decreased TSHβ mRNA stability.

    Original languageEnglish (US)
    Pages (from-to)469-475
    Number of pages7
    JournalMolecular Endocrinology
    Volume5
    Issue number4
    StatePublished - Apr 1991

    Fingerprint

    Poly A
    Thyrotropin
    Thyroid Hormones
    Messenger RNA
    RNA Stability
    RNA
    Half-Life
    Cyclophilin A
    Alternative Splicing
    Dactinomycin
    Triiodothyronine
    Cycloheximide
    Thyroxine

    ASJC Scopus subject areas

    • Molecular Biology
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Thyroid hormone decreases the stability and the poly(A) tract length of rat thyrotropin β-subunit messenger RNA. / Krane, Ian M.; Spindel, Eliot; Chin, William W.

    In: Molecular Endocrinology, Vol. 5, No. 4, 04.1991, p. 469-475.

    Research output: Contribution to journalArticle

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    N2 - Thyroid hormone (T3 and T4) down-regulation of TSH subunit steady state mRNA levels and subunit gene transcription in vitro and in vivo has been well studied. We present evidence here that T3 can also regulate the turnover of TSH subunit mRNA. The apparent half-life of the TSH β-subunit mRNA was determined by adding actinomycin-D (2 μM) to dispersed rat pituitary cultures in hypothyroid medium or medium containing 10-7 M T3 and analyzing the decline in subunit mRNA levels with time. The half-life of the TSHβ mRNA from those cultures treated with T3 was shorter than that of the control cultures (9 vs. >24 h, respectively). A possible mechanism by which TSH β-subunit mRNA stability is altered is through a change in the size of each mRNA's poly(A) tail. Northern blot analysis of total RNA from the above cultures revealed that T3 treatment reduces the size of the TSH β-subunit mRNA. To determine if this alteration of mRNA size was due to a loss of a portion of the poly(A) tract and not to alternative splicing of the transcript or use of a secondary transcriptional start site, pooled RNAs were hybridized with oligo(dT) and subsequently digested with RNAse-H to remove the poly(A) tract. RNA blot analysis of these RNAs showed that T3 treatment results in the loss of most of the TSHβ poly(A) tail. After 8 h in culture, the T3-induced shortening of the poly(A) tail is specific to the TSH β-subunit, as we saw no such effect on the similarly sized mRNAs (α-subunit, LHβ, or cyclophilin-A). The effect of T3 to shorten the TSHβ mRNA can be mimicked by cycloheximide (20 μg/ml) or actinomycin-D, indicating that a rapidly turning-over protein may be involved in maintaining the size and possibly the stability of the TSHβ mRNA under euthyroid and hypothyroid conditions. We have also observed that this T3-induced shortening of the TSHβ mRNA can be produced in vivo within 4 h after a single dose of T3 has been administered to euthyroid animals (20 μg/100 g BW, ip). Thus, T3 reduces the apparent half-life of the TSHβ mRNA and causes a concomitant decrease in its poly(A) tail size. Although a causal relationship has not been established, it is possible that shortening of the poly(A) tract may lead to decreased TSHβ mRNA stability.

    AB - Thyroid hormone (T3 and T4) down-regulation of TSH subunit steady state mRNA levels and subunit gene transcription in vitro and in vivo has been well studied. We present evidence here that T3 can also regulate the turnover of TSH subunit mRNA. The apparent half-life of the TSH β-subunit mRNA was determined by adding actinomycin-D (2 μM) to dispersed rat pituitary cultures in hypothyroid medium or medium containing 10-7 M T3 and analyzing the decline in subunit mRNA levels with time. The half-life of the TSHβ mRNA from those cultures treated with T3 was shorter than that of the control cultures (9 vs. >24 h, respectively). A possible mechanism by which TSH β-subunit mRNA stability is altered is through a change in the size of each mRNA's poly(A) tail. Northern blot analysis of total RNA from the above cultures revealed that T3 treatment reduces the size of the TSH β-subunit mRNA. To determine if this alteration of mRNA size was due to a loss of a portion of the poly(A) tract and not to alternative splicing of the transcript or use of a secondary transcriptional start site, pooled RNAs were hybridized with oligo(dT) and subsequently digested with RNAse-H to remove the poly(A) tract. RNA blot analysis of these RNAs showed that T3 treatment results in the loss of most of the TSHβ poly(A) tail. After 8 h in culture, the T3-induced shortening of the poly(A) tail is specific to the TSH β-subunit, as we saw no such effect on the similarly sized mRNAs (α-subunit, LHβ, or cyclophilin-A). The effect of T3 to shorten the TSHβ mRNA can be mimicked by cycloheximide (20 μg/ml) or actinomycin-D, indicating that a rapidly turning-over protein may be involved in maintaining the size and possibly the stability of the TSHβ mRNA under euthyroid and hypothyroid conditions. We have also observed that this T3-induced shortening of the TSHβ mRNA can be produced in vivo within 4 h after a single dose of T3 has been administered to euthyroid animals (20 μg/100 g BW, ip). Thus, T3 reduces the apparent half-life of the TSHβ mRNA and causes a concomitant decrease in its poly(A) tail size. Although a causal relationship has not been established, it is possible that shortening of the poly(A) tract may lead to decreased TSHβ mRNA stability.

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