Wreden C, Verrotti A C, Schisa J A, Lieberfarb M E, Strickland S
Department of Pharmacology, University Medical Center at Stony Brook, NY 11794-8651, USA.
Development. 1997 Aug;124(15):3015-23. doi: 10.1242/dev.124.15.3015.
Nanos protein promotes abdominal structures in Drosophila embryos by repressing the translation of maternal hunchback mRNA in the posterior. To study the mechanism of nanos-mediated translational repression, we first examined the mechanism by which maternal hunchback mRNA is translationally activated. In the absence of nanos activity, the poly(A) tail of hunchback mRNA is elongated concomitant with its translation, suggesting that cytoplasmic polyadenylation directs activation. However, in the presence of nanos the length of the hunchback mRNA poly(A) tail is reduced. To determine if nanos activity represses translation by altering the polyadenylation state of hunchback mRNA, we injected various in vitro transcribed RNAs into Drosophila embryos and determined changes in polyadenylation. Nanos activity reduced the polyadenylation status of injected hunchback RNAs by accelerating their deadenylation. Pumilio activity, which is necessary to repress the translation of hunchback, is also needed to alter polyadenylation. An examination of translation indicates a strong correlation between poly(A) shortening and suppression of translation. These data indicate that nanos and pumilio determine posterior morphology by promoting the deadenylation of maternal hunchback mRNA, thereby repressing its translation.
纳米蛋白通过抑制果蝇胚胎后部母体驼背mRNA的翻译来促进腹部结构的形成。为了研究纳米介导的翻译抑制机制,我们首先研究了母体驼背mRNA翻译激活的机制。在没有纳米活性的情况下,驼背mRNA的聚腺苷酸尾巴在其翻译过程中同时延长,这表明细胞质聚腺苷酸化指导激活。然而,在有纳米的情况下,驼背mRNA聚腺苷酸尾巴的长度会缩短。为了确定纳米活性是否通过改变驼背mRNA的聚腺苷酸化状态来抑制翻译,我们将各种体外转录的RNA注射到果蝇胚胎中,并确定聚腺苷酸化的变化。纳米活性通过加速其去腺苷酸化来降低注射的驼背RNA的聚腺苷酸化状态。抑制驼背翻译所必需的Pumilio活性对于改变聚腺苷酸化也是必需的。对翻译的检查表明聚(A)缩短与翻译抑制之间存在很强的相关性。这些数据表明,纳米和Pumilio通过促进母体驼背mRNA的去腺苷酸化来决定后部形态,从而抑制其翻译。