Avedisov S N, Il'in Iu V
Mol Biol (Mosk). 1994 Jul-Aug;28(4):813-21.
The presence of a third long open reading frame (ORF3) is the common feature of a number of Drosophila retrotransposons, including MDG4 (gypsy). Thus, these elements have a strong structural resemblance to the integrated forms of vertebrate retroviruses. To elucidate the mode of expression of ORF3, transcription analysis of MDG4 for several D. melanogaster strains and Schneider 2 cultured cells was carried out. In all cases the analysis revealed the presence of 2-kb subgenomic ORF3 transcripts in the polyadenylated RNA fractions. It was shown by using the cDNA-PCR technique with different sets of primers that these subgenomic 2-kb RNAs are generated through splicing of full-length MDG4 transcripts. In all cases, only one DNA fragment is amplified for each suitable pair of primers, indicating that there is probable no alternative splicing during the ORF3 expression. The primary structure of spliced RNA was determined, and its analysis shows that both MDG4 subfamilies (6K and 7K) are apparently able to encode functionally active translation products of ORF3. The regulation at the level of splicing is supposed to be one of the most important factors controlling the transposition frequency of MDG4.
第三个长开放阅读框(ORF3)的存在是包括MDG4(吉普赛)在内的许多果蝇逆转录转座子的共同特征。因此,这些元件在结构上与脊椎动物逆转录病毒的整合形式有很强的相似性。为了阐明ORF3的表达模式,对几种黑腹果蝇品系和施耐德2培养细胞进行了MDG4的转录分析。在所有情况下,分析都揭示了在多聚腺苷酸化RNA组分中存在2 kb的亚基因组ORF3转录本。通过使用不同引物组的cDNA-PCR技术表明,这些亚基因组2 kb RNA是通过全长MDG4转录本的剪接产生的。在所有情况下,对于每对合适的引物只扩增出一个DNA片段,这表明在ORF3表达过程中可能没有可变剪接。确定了剪接RNA的一级结构,分析表明MDG4的两个亚家族(6K和7K)显然都能够编码功能活性的ORF3翻译产物。剪接水平的调控被认为是控制MDG4转座频率的最重要因素之一。