Baeyens D A, Cornett L E
Department of Biology, University of Arkansas at Little Rock 72204, USA.
J Cell Physiol. 1995 May;163(2):305-11. doi: 10.1002/jcp.1041630211.
In the liver, transcript destabilization contributes to the decrease in steady-state levels of beta 2-adrenergic receptor mRNA that occurs during early postnatal development in the rat. From genomic DNA, polymerase chain reaction (PCR) was used to amplify a 718-basepair (bp) fragment of the beta 2-adrenergic receptor gene including the entire 3'-untranslated region. Results from SDS-gel electrophoresis and autoradiography demonstrated a M(r) 85,000 cellular factor present in postnatal day 60, but not fetal day 18 rat liver that was ultraviolet (UV) light-crosslinked to in vitro transcribed beta 2-adrenergic receptor RNA 3'-untranslated region. Unlabeled beta 2-adrenergic receptor RNA 3'-untranslated region, but not mouse beta-actin RNA, competed with labeled beta 2-adrenergic receptor RNA 3'-untranslated region for binding to the M(r) 85,000 protein. Cross-linking of the beta 2-adrenergic receptor RNA 3'-untranslated region to the M(r) 85,000 protein was inhibited by the ribohomopolymer poly(U), with poly(A), poly(C) and poly(G) having little or no effect. Thus, a M(r) 85,000 protein has been identified in adult male rat liver that may interact with U-rich sequences in the 3'-untranslated region of the beta 2-adrenergic receptor mRNA and may account for the decreased stability of hepatic beta 2-adrenergic receptor gene transcripts that occurs during development.
在肝脏中,转录本去稳定化导致大鼠出生后早期发育过程中β2 - 肾上腺素能受体mRNA稳态水平的降低。利用聚合酶链反应(PCR)从基因组DNA中扩增出一段718个碱基对(bp)的β2 - 肾上腺素能受体基因片段,该片段包括整个3' - 非翻译区。十二烷基硫酸钠 - 凝胶电泳(SDS - gel electrophoresis)和放射自显影结果表明,在出生后第60天的大鼠肝脏中存在一种分子量为85,000的细胞因子,但在胚胎第18天的大鼠肝脏中不存在,该细胞因子可与体外转录的β2 - 肾上腺素能受体RNA 3' - 非翻译区发生紫外线(UV)光交联。未标记的β2 - 肾上腺素能受体RNA 3' - 非翻译区,而非小鼠β - 肌动蛋白RNA,能与标记的β2 - 肾上腺素能受体RNA 3' - 非翻译区竞争结合分子量为85,000的蛋白质。β2 - 肾上腺素能受体RNA 3' - 非翻译区与分子量为85,000的蛋白质的交联被核糖同聚物聚(U)抑制,而聚(A)、聚(C)和聚(G)的影响很小或没有影响。因此,在成年雄性大鼠肝脏中鉴定出一种分子量为85,000的蛋白质,它可能与β2 - 肾上腺素能受体mRNA 3' - 非翻译区富含U的序列相互作用,并可能解释发育过程中肝脏β2 - 肾上腺素能受体基因转录本稳定性降低的原因。