Boothby M, Kukowska J, Boime I
J Biol Chem. 1983 Aug 10;258(15):9250-3.
Previous studies have demonstrated an imbalance in placental levels of the human choriogonadotropin (hCG) alpha and beta subunits. Free alpha subunit was present in first trimester placentae, and the imbalance was accentuated as gestation approached parturition. Two sets of experiments were performed to assess the control on production levels of each subunit. Synthesis of the alpha and beta subunits was assessed by labeling the nascent chains of polysomes derived from first trimester placenta. The products of these reactions were immunoprecipitated with subunit-specific antisera and the labeled subunits were quantitated; the ratio of alpha to beta subunit synthesized was 1.7. To examine whether this imbalanced synthesis reflected differences in the amount of subunit mRNAs, or differing mRNA translational efficiencies, the ratio of the steady state levels of these mRNAs was also determined. Total first trimester placental RNA was hydrolyzed with alkali, 5'-end-labeled with 32P, and hybridized in DNA excess to cloned alpha and beta cDNAs. These experiments demonstrated the presence of twice as much hCG-alpha mRNA as hCG-beta mRNA. In term placenta, the amounts of excess alpha subunit are greater than at first trimester; the ratio of alpha to beta mRNAs in term RNA was about 12:1. Thus, the subunit mRNA levels are independently regulated and their imbalance accounts for differences in the quantities of alpha and beta subunits seen in placental tissue.
先前的研究表明,人绒毛膜促性腺激素(hCG)α和β亚基在胎盘水平上存在失衡。游离α亚基存在于孕早期胎盘,随着妊娠接近分娩,这种失衡加剧。进行了两组实验来评估对每个亚基产生水平的控制。通过标记来自孕早期胎盘的多聚核糖体的新生链来评估α和β亚基的合成。这些反应的产物用亚基特异性抗血清进行免疫沉淀,并对标记的亚基进行定量;合成的α与β亚基的比例为1.7。为了研究这种合成失衡是否反映了亚基mRNA量的差异,或不同的mRNA翻译效率,还测定了这些mRNA稳态水平的比例。用碱水解孕早期胎盘总RNA,用32P进行5'-末端标记,并在DNA过量的情况下与克隆的α和β cDNA杂交。这些实验表明,hCG-α mRNA的量是hCG-β mRNA的两倍。在足月胎盘中,过量α亚基的量比孕早期更大;足月RNA中α与β mRNA的比例约为12:1。因此,亚基mRNA水平是独立调节的,它们的失衡解释了胎盘组织中α和β亚基数量的差异。