Searles L L, Ruth R S, Pret A M, Fridell R A, Ali A J
Department of Biology, University of North Carolina, Chapel Hill 27599-3280.
Mol Cell Biol. 1990 Apr;10(4):1423-31. doi: 10.1128/mcb.10.4.1423-1431.1990.
The nucleotide sequence and intron-exon structure of the Drosophila melanogaster vermilion (v) gene have been determined. In addition, the sites of several mutations and the effects of these mutations on transcription have been examined. The major v mRNA is generated upon splicing six exons of lengths (5' to 3') 83, 161, 134, 607, 94, and 227 nucleotides (nt). A minor species of v mRNA is initiated at an upstream site and has a 5' exon of at least 152 nt which overlaps the region included in the 83-nt exon of the major v RNA. The three v mutations, v1, v2, and vk, which can be suppressed by mutations at suppressor of sable, su(s), are insertions of transposon 412 at the same position in exon 1, 36 nt downstream of the major transcription initiation site. Despite the 7.5-kilobase insertion in these v alleles, a reduced level of wild-type-sized mRNA accumulates in suppressed mutant strains. The structure and transcription of several unsuppressible v alleles have also been examined. The v36f mutation is a B104/roo insertion in intron 4 near the splice donor site. A mutant carrying this alteration accumulates a very low level of mRNA that is apparently polyadenylated at a site within the B104/roo transposon. The v48a mutation, which deletes approximately 200 nt of DNA, fuses portions of exons 3 and 4 without disruption of the translational reading frame. A smaller transcript accumulates at a wild-type level, and thus an altered, nonfunctional polypeptide is likely to be synthesized in strains carrying this mutation. The v(H2a) mutants has a P element insertion in exon 6 within the coding region.
已确定黑腹果蝇朱红色(v)基因的核苷酸序列和内含子-外显子结构。此外,还研究了几个突变位点以及这些突变对转录的影响。主要的v mRNA是通过拼接六个长度(从5'到3')分别为83、161、134、607、94和227个核苷酸(nt)的外显子产生的。一种次要的v mRNA物种起始于上游位点,其5'外显子至少有152 nt,与主要v RNA的83-nt外显子所包含的区域重叠。三种v突变,即v1、v2和vk,可被黑貂抑制子(su(s))的突变所抑制,它们是转座子412插入到外显子1的同一位置,位于主要转录起始位点下游36 nt处。尽管这些v等位基因中有7.5千碱基的插入,但在被抑制的突变菌株中仍积累了野生型大小的mRNA,不过水平有所降低。还研究了几个不可抑制的v等位基因的结构和转录情况。v36f突变是B104/roo插入到内含子4中靠近剪接供体位点的位置。携带这种改变的突变体积累的mRNA水平非常低,显然是在B104/roo转座子内的一个位点进行了多聚腺苷酸化。v48a突变缺失了大约200 nt的DNA,将外显子3和4的部分融合在一起,而没有破坏翻译阅读框。一种较小的转录本以野生型水平积累,因此在携带这种突变的菌株中可能合成了一种改变的、无功能的多肽。v(H2a)突变体在编码区内的外显子6中有一个P元件插入。