Cherbas L, Schulz R A, Koehler M M, Savakis C, Cherbas P
J Mol Biol. 1986 Jun 20;189(4):617-31. doi: 10.1016/0022-2836(86)90492-4.
The EIPs 28 and 29 are a family of polypeptides identified originally by their ecdysone inducibility in Drosophila cell lines. At least two family members, 28III and 29III, appear to be primary translation products. Here we describe a unique Eip28/29 gene that must encode both primary products. The Eip28/29 gene is unique because the cloned genomic DNA hybridizes to both EIP 28 and 29 messenger RNAs under stringent conditions, but does not anneal detectably to other genomic sequences even under mild conditions. Furthermore the diverse products of this gene are not alleles because flies homozygous for the chromosomal region (71CD) containing the Eip28/29 gene produce mRNAs that translate to yield all the EIPs 28 and 29. We report here the sequence of a 2855-nucleotide region encompassing the Eip28/29 gene. By comparisons with complementary DNA sequences and by nuclease protection experiments we have derived a complete structure for the Eip28/29 transcription unit. The primary transcript is 2146 nucleotides long and is processed by the removal of three introns to yield the predominant mature transcript in tissue culture cells (979 nucleotides). This transcript probably corresponds to the 28III mRNA. Neither the start of the transcription unit nor the structure of the predominant transcript is affected by the hormone ecdysone. The genomic sequence reveals a series of heptanucleotide and octanucleotide repeats of unknown function that fall at about 50-nucleotide intervals within the first 150 nucleotides upstream from the transcription unit. In addition this sequence, when combined with previously published data, suggests that the consensus cap site sequence in Drosophila may be extended to include 13 nucleotides centered on the heptanucleotide core previously recognized by Snyder et al. (1982).
EIP28和EIP29是一类最初在果蝇细胞系中因其对蜕皮激素的诱导性而被鉴定出的多肽家族。至少两个家族成员,即28III和29III,似乎是初级翻译产物。在此,我们描述了一个独特的Eip28/29基因,它必定编码这两种初级产物。Eip28/29基因之所以独特,是因为克隆的基因组DNA在严格条件下能与EIP28和EIP29信使RNA杂交,但即使在温和条件下也未检测到与其他基因组序列退火。此外,该基因的多种产物并非等位基因,因为对于包含Eip28/29基因的染色体区域(71CD)纯合的果蝇会产生能翻译出所有EIP28和EIP29的信使RNA。我们在此报告了一个包含Eip28/29基因的2855个核苷酸区域的序列。通过与互补DNA序列比较以及核酸酶保护实验,我们推导得出了Eip28/29转录单位的完整结构。初级转录本长度为2146个核苷酸,通过去除三个内含子进行加工,从而在组织培养细胞中产生主要的成熟转录本(979个核苷酸)。该转录本可能对应于28III信使RNA。转录单位的起始点以及主要转录本的结构均不受蜕皮激素的影响。基因组序列揭示了一系列功能未知的七核苷酸和八核苷酸重复序列,它们在转录单位上游前150个核苷酸内以约50个核苷酸的间隔分布。此外,该序列与先前发表的数据相结合表明,果蝇中的共有帽位点序列可能会扩展,以包括以Snyder等人(1982年)先前识别的七核苷酸核心为中心的13个核苷酸。