Europe-Finner G N, Phaneuf S, Mardon H J, López Bernal A
University of Oxford, Nuffield Department of Obstetrics and Gynecology, John Radcliffe Hospital, Headington, United Kingdom.
J Clin Endocrinol Metab. 1996 Mar;81(3):1069-75. doi: 10.1210/jcem.81.3.8772578.
In previous studies using specific G alpha s antibodies we have identified several human myometrial G alpha s protein isoforms with molecular masses of 45, 46, 47, 54, and 58 kDa, respectively. During pregnancy, levels of the 46- and 54-kDa proteins are significantly increased compared to those in nonpregnant myometrium and then decreased at the onset of labor. In this study we investigated the expression of G alpha s messenger ribonucleic acid (mRNA) splice variants, which are generated as a result of alternative splicing of a single mRNA precursor, in term pregnancy and parturition to determine whether there was any correlation with the observed changes in G alpha s protein isoforms. A myometrial G alpha s complementary DNA was synthesized using RT-PCR and cloned into pCRtmII suitable for preparation of riboprobes for use in ribonuclease protection assays. Using this technique, we identified at least three myometrial G alpha s mRNAs, including two forms of G alpha s-Large (with or without the serine at amino acid 87) and one form of G alpha s-Small (with the serine at amino acid 72). G alpha s Small (with the serine) and G alpha s-Large (with the serine) mRNAs encode for the 46- and 54-kDa G alpha s protein isoforms, respectively, and were increased in term pregnancy and then subsequently decreased after the onset of labor. Our data suggest that posttranscriptional regulation of G alpha s mRNAs may be important in the differential expression of G alpha s protein isoforms during pregnancy and labor.
在之前使用特异性Gαs抗体的研究中,我们鉴定出了几种人子宫肌层Gαs蛋白亚型,其分子量分别为45、46、47、54和58 kDa。在怀孕期间,与非妊娠子宫肌层相比,46 kDa和54 kDa蛋白的水平显著升高,然后在分娩开始时下降。在本研究中,我们调查了Gαs信使核糖核酸(mRNA)剪接变体的表达情况,这些变体是由单个mRNA前体的可变剪接产生的,在足月妊娠和分娩过程中,以确定其与观察到的Gαs蛋白亚型变化是否存在任何相关性。使用逆转录聚合酶链反应(RT-PCR)合成子宫肌层Gαs互补DNA,并将其克隆到适合制备用于核糖核酸酶保护分析的核糖探针的pCRtmII中。使用该技术,我们鉴定出至少三种子宫肌层Gαs mRNA,包括两种形式的Gαs-Large(氨基酸87处有或没有丝氨酸)和一种形式的Gαs-Small(氨基酸72处有丝氨酸)。Gαs Small(有丝氨酸)和Gαs-Large(有丝氨酸)mRNA分别编码46 kDa和54 kDa的Gαs蛋白亚型,在足月妊娠时增加,然后在分娩开始后随后下降。我们的数据表明,Gαs mRNA的转录后调控可能在妊娠和分娩期间Gαs蛋白亚型的差异表达中起重要作用。