Autoregulation at the level of mRNA 3′ end formation of the suppressor of forked gene of Drosophila melanogaster is conserved in Drosophila virilis
Autoregulation at the level of mRNA 3′ end formation of the suppressor of forked gene of Drosophila melanogaster is conserved in Drosophila virilis
复制标题
DOI：
10.1073/pnas.95.24.14302
复制
发表时间：
1998-11-24
影响因子：
11.1
通讯作者：
Simonelig, M
中科院分区：
文献类型：
--
作者：
Audibert, A;Simonelig, M
果蝇叉形 [Su(f)] 蛋白的抑制蛋白与酵母 RNA14 蛋白和人裂解刺激因子的 77-kDa 亚基具有同源性，这些蛋白是参与 mRNA 3' 末端形成的蛋白。这表明 Su(f) 在果蝇 mRNA 3' 末端形成中发挥作用。 su(f) 基因产生三个转录本；其中两个在转录单元末端进行聚腺苷酸化，另一种是截短转录本，在内含子 4 中进行聚腺苷酸化。使用温度敏感的 su(f) 突变体，我们表明截短转录本的积累需要野生型 Su(f) 蛋白。这表明 Su(f) 蛋白通过刺激截短的 su(f) RNA 的 3' 端形成来负向自动调节其积累。从果蝇 virilis 中克隆 su(f) 并对其 RNA 谱进行分析表明，su(f) 的自动调节在该物种中是保守的。两个物种的 su(f) 之间的序列比较使我们能够指出截短的 RNA poly(A) 位点下游的内含子 4 中的三个保守区域。这些保守区域包括参与 Poly(A) 位点定义的富含 GU 的下游序列。使用内含子 4 内截短的转基因，我们表明直到富含 GU 的保守结构域的序列足以产生截短的 RNA 并足以通过 su(f) 调节这种产生。我们的结果表明su(f)在调节poly(A)位点利用中的作用以及富含GU的序列对于这种调节发生的重要作用。
The Drosophila melanogaster Suppressor of forked [Su(f)] protein shares homology with the yeast RNA14 protein and the 77-kDa subunit of human cleavage stimulation factor, which are proteins involved in mRNA 3' end formation. This suggests a role for Su(f) in mRNA 3' end formation in Drosophila. The su(f) gene produces three transcripts; two of them are polyadenylated at the end of the transcription unit, and one is a truncated transcript, polyadenylated in intron 4. Using temperature sensitive su(f) mutants, we show that accumulation of the truncated transcript requires wild-type Su(f) protein. This suggests that the Su(f) protein autoregulates negatively its accumulation by stimulating 3' end formation of the truncated su(f) RNA. Cloning of su(f) from Drosophila virilis and analysis of its RNA profile suggest that su(f) autoregulation is conserved in this species. Sequence comparison between su(f) from both species allows us to point out three conserved regions in intron 4 downstream of the truncated RNA poly(A) site. These conserved regions include the GU-rich downstream sequence involved in poly(A) site definition. Using transgenes truncated within intron 4, we show that sequence up to the conserved GU-rich domain is sufficient for production of the truncated RNA and for regulation of this production by su(f). Our results indicate a role of su(f) in the regulation of poly(A) site utilization and an important role of the GU-rich sequence for this regulation to occur.