Davis R L, Davidson N
Mol Cell Biol. 1986 May;6(5):1464-70. doi: 10.1128/mcb.6.5.1464-1470.1986.
We have previously isolated the region of the Drosophila melanogaster X chromosome which contains the dunce+ gene and mapped the dunce2 mutation by recombination to a 10- to 12-kilobase (kb) interval (R. L. Davis and N. Davidson, Mol. Cell. Biol. 4:358-367, 1984). Here, we examine the expression of the dunce+ chromosomal region and identify the dunce+ gene within that region. A region of ca. 25 kb which contains the 10- to 12-kb interval to which dunce2 was mapped codes for polyadenylated RNAs of 9.6, 7.4, 7.2, 7.0, 5.4, and 4.5 kb in adult flies. These transcripts are encoded by the same DNA strand and share sequences of some exons, indicating that the transcripts arise from the same gene. Some genome probes internal to the ca. 25-kb coding region show transcript-specific hybridization, demonstrating alternate usage of exonic sequence information in the formation of the mature transcripts. The basis for this internal heterogeneity in RNAs is most likely alternative splicing. Two dunce mutants examined show aberrant RNA expression from this coding region, confirming that this region is the dunce gene. The developmental expression of these transcripts has been examined. The 5.4-kb RNA is present at all developmental stages. The 9.6-, 7.4-, 7.2-, and 7.0-kb RNAs are not expressed at detectable levels in embryos, but are detected in late embryogenesis and in later developmental stages. The 4.5-kb species is found in early embryos and adults, but not in intermediate stages. We discuss the remarkable transcript heterogeneity and expression pattern with respect to the important function this gene performs in neurobiological and other physiological processes.
我们之前分离出了果蝇黑腹果蝇X染色体上包含dunce+基因的区域,并通过重组将dunce2突变定位到了一个10至12千碱基(kb)的区间(R. L. 戴维斯和N. 戴维森,《分子细胞生物学》4:358 - 367,1984年)。在此,我们研究了dunce+染色体区域的表达情况,并在该区域内鉴定出了dunce+基因。一个约25 kb的区域,其中包含dunce2被定位到的10至12 kb区间,在成年果蝇中编码9.6、7.4、7.2、7.0、5.4和4.5 kb的多聚腺苷酸化RNA。这些转录本由同一条DNA链编码,并共享一些外显子序列,表明这些转录本来自同一个基因。约25 kb编码区域内的一些基因组探针显示出转录本特异性杂交,证明在成熟转录本形成过程中外显子序列信息存在交替使用的情况。RNA这种内部异质性的基础很可能是可变剪接。所检测的两个dunce突变体在这个编码区域表现出异常的RNA表达,证实该区域就是dunce基因。我们已经研究了这些转录本的发育表达情况。5.4 kb的RNA在所有发育阶段都存在。9.6、7.4、7.2和7.0 kb的RNA在胚胎中未检测到可检测水平的表达,但在胚胎后期发育和后续发育阶段可检测到。4.5 kb的种类在早期胚胎和成虫中存在,但在中间阶段不存在。我们讨论了这种显著的转录本异质性和表达模式与该基因在神经生物学和其他生理过程中所执行的重要功能的关系。