Coulson R M, Connor V, Chen J C, Ajioka J W
Department of Pathology, University of Cambridge, UK.
Mol Biochem Parasitol. 1996 Nov 25;82(2):227-36. doi: 10.1016/0166-6851(96)02739-9.
Tubulin expression has been analysed as the insect stage of the protozoan parasite Leishmania major differentiates from a non-infective to an infective form. This transformation of the promastigote stage occurs in vitro and analysis of beta-tubulin mRNA expression in axenically grown promastigotes showed that a 2200 nt transcript is predominately expressed in non-infective promastigotes. The message contains a motif associated with mRNA intracellular localisation and its level is reduced by an order of magnitude in infective promastigotes through a mechanism involving RNA stability. A 3200 nt RNA, the major beta-tubulin transcript in the infective stage, is encoded by a single copy gene at the 3' end of the array that encodes the 2200 nt RNA. These RNAs, as well as a gene encoding a beta-tubulin transcript highly up-regulated in the mammalian stage of the parasite, encode polypeptides that are apparently functionally equivalent but have highly diverged 3' untranslated regions. This differential regulation of the dispersed isogenes may reflect the involvement of a mechanism altering tubulin synthesis during the Leishmania life cycle. The analysis of alpha-tubulin RNA levels revealed the abundance of this message falls as promastigotes differentiate into an infectious stage and the transcript is destabilised in infective promastigotes. These data demonstrate that the regulation of mRNA half-life contributes to controlling gene expression as promastigotes differentiate into an infectious form.
随着原生动物寄生虫硕大利什曼原虫的昆虫阶段从不感染形式分化为感染形式,微管蛋白的表达已得到分析。前鞭毛体阶段的这种转变在体外发生,对在无菌培养的前鞭毛体中β-微管蛋白mRNA表达的分析表明,一种2200 nt的转录本在非感染性前鞭毛体中大量表达。该信息包含一个与mRNA细胞内定位相关的基序,并且通过涉及RNA稳定性的机制,其水平在感染性前鞭毛体中降低了一个数量级。一种3200 nt的RNA,即感染阶段的主要β-微管蛋白转录本,由编码2200 nt RNA的阵列3'端的一个单拷贝基因编码。这些RNA,以及一个在寄生虫的哺乳动物阶段高度上调的编码β-微管蛋白转录本的基因,编码的多肽在功能上显然是等效的,但具有高度分化的3'非翻译区。这些分散的同基因的差异调节可能反映了利什曼原虫生命周期中改变微管蛋白合成的机制的参与。α-微管蛋白RNA水平的分析表明,随着前鞭毛体分化为感染阶段,该信息的丰度下降,并且转录本在感染性前鞭毛体中不稳定。这些数据表明,mRNA半衰期的调节有助于控制前鞭毛体分化为感染形式时的基因表达。