Georgieva T, Dunkov B C, Harizanova N, Ralchev K, Law J H
Department of Biochemistry and Center for Insect Science, University of Arizona, Tucson, AZ 85721, USA.
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2716-21. doi: 10.1073/pnas.96.6.2716.
Insect ferritins have subunits homologous to the heavy and light chains of vertebrate ferritins. Cloning and sequence of the heavy chain homologue (HCH) of Drosophila melanogaster ferritin subunit have been reported earlier. When Northern blots of D. melanogaster RNA were probed with a cDNA for this HCH, three bands were observed. It was shown that these represented at least four classes of mRNA of various lengths. The polymorphism results from alternative splicing of an intron in the 5' untranslated region (UTR) that contains the iron-responsive element (IRE) and from two alternative polyadenylation sites in the 3' UTR. This has also been reported by others [Lind, M. I., Ekengren, S., Melefors, O. & Söderhäll, K. (1998) FEBS Lett. 436, 476-482]. By hybridizing Northern blots with specific probes, it has been shown that the relative proportions of the messages vary with the life stage and especially with iron supplementation of the diet. Iron significantly increases the amount of ferritin HCH messages and dramatically shifts the balance toward those messages that lack an IRE and/or have a short 3' UTR. In the larvae this change takes place in the gut, but not in the fat body. We speculate that this dramatic shift in message distribution may result from an effect of iron on the rate of transcription or message degradation, or from an effect on the splicing process itself. Synthesis of ferritin HCH subunit mRNAs that lack an IRE may be important under conditions of iron overload.
昆虫铁蛋白具有与脊椎动物铁蛋白重链和轻链同源的亚基。果蝇铁蛋白亚基重链同源物(HCH)的克隆和序列先前已有报道。当用该HCH的cDNA探测果蝇RNA的Northern印迹时,观察到三条带。结果表明,这些带至少代表了四种不同长度的mRNA类别。这种多态性源于5'非翻译区(UTR)中包含铁反应元件(IRE)的内含子的可变剪接以及3'UTR中的两个可变聚腺苷酸化位点。其他人也有过相关报道[林德,M.I.,埃肯格伦,S.,梅勒福斯,O.和索德哈尔,K.(1998年)《欧洲生物化学学会联合会快报》436,476 - 482]。通过用特异性探针与Northern印迹杂交,已表明这些信息的相对比例随生命阶段变化,尤其是随饮食中铁的补充而变化。铁显著增加铁蛋白HCH信息的量,并极大地使平衡向那些缺乏IRE和/或具有短3'UTR的信息转移。在幼虫中,这种变化发生在肠道而非脂肪体中。我们推测,这种信息分布的显著变化可能是由于铁对转录速率或信息降解的影响,或者是对剪接过程本身的影响。在铁过载的情况下,缺乏IRE的铁蛋白HCH亚基mRNA的合成可能很重要。