Department of Human Biochemistry, School of Medicine, Instituto de Investigaciones Moleculares de Enfermedades Hormonales Neurodegenerativas y Oncológicas, University of Buenos Aires, Buenos Aires, Argentina.
PLoS One. 2011;6(8):e22822. doi: 10.1371/journal.pone.0022822. Epub 2011 Aug 1.
Cholesterol transport is essential for many physiological processes, including steroidogenesis. In steroidogenic cells hormone-induced cholesterol transport is controlled by a protein complex that includes steroidogenic acute regulatory protein (StAR). Star is expressed as 3.5-, 2.8-, and 1.6-kb transcripts that differ only in their 3'-untranslated regions. Because these transcripts share the same promoter, mRNA stability may be involved in their differential regulation and expression. Recently, the identification of natural antisense transcripts (NATs) has added another level of regulation to eukaryotic gene expression. Here we identified a new NAT that is complementary to the spliced Star mRNA sequence. Using 5' and 3' RACE, strand-specific RT-PCR, and ribonuclease protection assays, we demonstrated that Star NAT is expressed in MA-10 Leydig cells and steroidogenic murine tissues. Furthermore, we established that human chorionic gonadotropin stimulates Star NAT expression via cAMP. Our results show that sense-antisense Star RNAs may be coordinately regulated since they are co-expressed in MA-10 cells. Overexpression of Star NAT had a differential effect on the expression of the different Star sense transcripts following cAMP stimulation. Meanwhile, the levels of StAR protein and progesterone production were downregulated in the presence of Star NAT. Our data identify antisense transcription as an additional mechanism involved in the regulation of steroid biosynthesis.
胆固醇运输对于许多生理过程都是必不可少的,包括类固醇生成。在类固醇生成细胞中,激素诱导的胆固醇运输受包括类固醇急性调节蛋白 (StAR) 在内的蛋白质复合物控制。Star 以 3.5、2.8 和 1.6kb 的转录本表达,仅在其 3'非翻译区存在差异。由于这些转录本共享相同的启动子,mRNA 稳定性可能参与它们的差异调节和表达。最近,天然反义转录本 (NAT) 的鉴定为真核基因表达增加了另一个调节层次。在这里,我们鉴定了一种与剪接 Star mRNA 序列互补的新 NAT。使用 5' 和 3' RACE、链特异性 RT-PCR 和核糖核酸酶保护测定,我们证明 Star NAT 在 MA-10 黄体细胞和类固醇生成的鼠组织中表达。此外,我们确定人绒毛膜促性腺激素通过 cAMP 刺激 Star NAT 表达。我们的结果表明,由于它们在 MA-10 细胞中共表达,因此 sense-antisense Star RNAs 可能会协调调节。cAMP 刺激后,Star NAT 的过表达对不同 Star sense 转录本的表达产生了不同的影响。同时,在存在 Star NAT 的情况下,StAR 蛋白水平和孕酮产生被下调。我们的数据表明,反义转录是参与类固醇生物合成调节的另一种机制。