Translational control by Trypanosoma brucei DRBD18 contributes to the maintenance of the procyclic state.
Translational control by Trypanosoma brucei DRBD18 contributes to the maintenance of the procyclic state.
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DOI：
10.1261/rna.079625.123
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发表时间：
2023-12
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影响因子：
--
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布氏锥虫在其整个生命周期中，在哺乳动物宿主和采采蝇宿主内都占据着不同的生态位。这些环境各自的免疫学和生物化学复杂性及多变性要求寄生虫重塑其蛋白质图谱，以逃避监视并应对不断变化的代谢需求。在包括布氏锥虫在内的动基体目原生动物中，转录后控制机制是基因调控的主要手段，且这些机制通常由RNA结合蛋白介导。DRBD18是一种布氏锥虫RNA结合蛋白，据报道它与核糖体蛋白和翻译因子相互作用。在此，我们测试了DRBD18在翻译控制中的作用。我们验证了DRBD18与正在翻译的核糖体以及翻译起始因子eIF3a的相互作用。我们进一步表明，通过RNA干扰使DRBD18缺失会导致多聚核糖体图谱发生改变，特别是重多聚核糖体减少。核糖体图谱分析显示，在DRBD18缺失时，有101种转录本的翻译效率（TE）发生变化：41种表现出TE降低，60种表现出TE升高。另外还有66种转录本得到缓冲，即转录本丰度的变化通过TE的变化得到补偿，使得总的翻译产出预计不会改变。在DRBD18缺失的细胞中，一组编码前循环型特异性蛋白的转录本在翻译上受到抑制，而相反，编码血流型和后循环型特异性蛋白的转录本在翻译上得到增强。RNA免疫沉淀/实时定量聚合酶链反应表明，DRBD18与受抑制和增强的转录本群体的成员都有关联。这些数据表明，DRBD18通过对特定mRNA进行正向和负向翻译控制，有助于维持前循环状态。
Trypanosoma brucei occupies distinct niches throughout its life cycle, within both the mammalian and tsetse fly hosts. The immunological and biochemical complexity and variability of each of these environments require a reshaping of the protein landscape of the parasite both to evade surveillance and face changing metabolic demands. In kinetoplastid protozoa, including T. brucei, posttranscriptional control mechanisms are the primary means of gene regulation, and these are often mediated by RNA-binding proteins. DRBD18 is a T. brucei RNA-binding protein that reportedly interacts with ribosomal proteins and translation factors. Here, we tested a role for DRBD18 in translational control. We validate the DRBD18 interaction with translating ribosomes and the translation initiation factor, eIF3a. We further show that DRBD18 depletion by RNA interference leads to altered polysomal profiles with a specific depletion of heavy polysomes. Ribosome profiling analysis reveals that 101 transcripts change in translational efficiency (TE) upon DRBD18 depletion: 41 exhibit decreased TE and 60 exhibit increased TE. A further 66 transcripts are buffered, that is, changes in transcript abundance are compensated by changes in TE such that the total translational output is expected not to change. In DRBD18-depleted cells, a set of transcripts that codes for procyclic form-specific proteins is translationally repressed while, conversely, transcripts that code for bloodstream form- and metacyclic form-specific proteins are translationally enhanced. RNA immunoprecipitation/qRT-PCR indicates that DRBD18 associates with members of both repressed and enhanced cohorts. These data suggest that DRBD18 contributes to the maintenance of the procyclic state through both positive and negative translational control of specific mRNAs.
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影响因子：
4.8
作者：
Chávez S;Urbaniak MD;Benz C;Smircich P;Garat B;Sotelo-Silveira JR;Duhagon MA
通讯作者：
Duhagon MA
影响因子：
3.7
作者：
Ciganda M;Prohaska K;Hellman K;Williams N
通讯作者：
Williams N
影响因子：
3.3
作者：
Durante IM;Butenko A;Rašková V;Charyyeva A;Svobodová M;Yurchenko V;Hashimi H;Lukeš J
通讯作者：
Lukeš J
影响因子：
6.7
作者：
McDonald L;Cayla M;Ivens A;Mony BM;MacGregor P;Silvester E;McWilliam K;Matthews KR
通讯作者：
Matthews KR
影响因子：
46.9
作者：
Fredriksson, S;Gullberg, M;Landegren, U
通讯作者：
Landegren, U