Komonyi O, Mink M, Csiha J, Maróy P
Department of Genetics, A.J. University, Szeged, Hungary.
Arch Insect Biochem Physiol. 1998;38(4):185-92. doi: 10.1002/(SICI)1520-6327(1998)38:4<185::AID-ARCH4>3.0.CO;2-Q.
The recombinant lambda clone 4-2 containing the genomic region of the Drosophila hormone receptor 38 (DHR 38) gene, homologous to mammalian neuronal growth factor I-B (NGFI-B), was isolated by radioactive labelled oligonucleotide hybridization. The nucleotide sequence of the genomic clone revealed three exons that encode the functional domains of the protein. The N-terminal exon1 had no homology at the amino acid level with NGFI-B, the mammalian homologue. A glutamine-rich region, probably involved in transcriptional activation, was observed at the C-terminal part of this exon. A similar motif is also present upstream in another reading frame of the same strand. Both motifs are preceded by a repetitive non-anucleotide sequence containing an AluI site, resembling a duplicated human Alu-sequence. A monomeric version of this sequence, coding similarly for an oligoglutamine peptide, occurs at a surprisingly high frequency in other regulatory genes in Drosophila. In contrast to mammalian Alu sequences, this sequence is found almost exclusively in the coding regions of Drosophila genes, but not in the non-coding parts of the genes. The DNA-binding domain with two zinc-fingers, and at least part of the ligand-binding peptide, is coded by the largest middle exon2 in DHR38 and exhibits up to 100% homology in short peptide motifs to its mammalian counterpart, where these domains are split into exons 3, 4, 5, and 6. However, the length, information content, stop codon, and splice site are conserved in the last exons in both fly and rat. In situ hybridization to 0-24 h wholemount embryos showed strong expression of DHR38 in neurogenic regions and in the intestinal tract during embryogenesis, suggesting a spatial and temporal control of transcription, partially analogous to the central nervous system-specific expression of NGFI-B in mammals.
通过放射性标记的寡核苷酸杂交,分离出了重组λ克隆4-2,它包含果蝇激素受体38(DHR 38)基因的基因组区域,该基因与哺乳动物神经生长因子I-B(NGFI-B)同源。基因组克隆的核苷酸序列显示有三个外显子,它们编码该蛋白质的功能结构域。N端的外显子1在氨基酸水平上与哺乳动物同源物NGFI-B没有同源性。在该外显子的C端部分观察到一个富含谷氨酰胺的区域,可能参与转录激活。在同一条链的另一个阅读框的上游也存在类似的基序。这两个基序之前都有一个包含AluI位点的重复非九核苷酸序列,类似于重复的人类Alu序列。该序列的单体形式,同样编码一个寡聚谷氨酰胺肽,在果蝇的其他调控基因中出现的频率出奇地高。与哺乳动物的Alu序列不同,该序列几乎只存在于果蝇基因的编码区域,而不存在于基因的非编码部分。具有两个锌指的DNA结合结构域以及至少部分配体结合肽,由DHR38中最大的中间外显子2编码,并且在短肽基序上与其哺乳动物对应物具有高达100%的同源性,在哺乳动物中这些结构域被分成外显子3、4、5和6。然而,果蝇和大鼠最后一个外显子中的长度、信息含量、终止密码子和剪接位点是保守的。对0 - 24小时的整装胚胎进行原位杂交显示,在胚胎发育过程中,DHR38在神经发生区域和肠道中有强烈表达,这表明转录存在时空控制,部分类似于哺乳动物中NGFI-B在中枢神经系统中的特异性表达。