de Brito Tarcísio Fontenele, Arruda Cardoso Maira, Atinbayeva Nazerke, Alexandre de Abreu Brito Ingrid, Amaro da Costa Lucas, Iovino Nicola, Pane Attilio
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Department of Chromatin Regulation, Max Planck Institute of Immunobiology and Epigenetics, Freiburg im Breisgau, Germany.
Front Cell Dev Biol. 2024 Nov 20;12:1481881. doi: 10.3389/fcell.2024.1481881. eCollection 2024.
Piwi proteins and the associated Piwi-interacting RNAs (piRNAs) coordinate a surveillance system that protects the animal genome from DNA damage induced by transposable element (TE) mobilization. While the pathway has been described in detail in the fruit fly , much less is known in more basal insects. is an hemipteran insect and one of the major vectors of Chagas disease. acquired specific classes of horizontally transferred transposons (HTTs) by feeding on bats, opossums and squirrel monkeys, thus providing the opportunity to investigate the piRNA-base response against HTTs in this species.
SmallRNA-Seq reads mapping to HTTs and resident transposable elements were quantified and checked for piRNA features like 1U a 10A biases, ping-pong and phasing signatures. Uniquely mapped piRNAs were used to identify piRNA clusters in ' genome. RNA-Seq data was used to quantify transposon and Rp-PIWI genes expression levels and were validated by qRT-PCR.
By analyzing the temporal dynamics of piRNA cluster expression and piRNA production during critical stages of Rhodnius development, we show that peak levels of ∼28 nt long piRNAs correlate with reduced HTT and resident TE expression primarily during embryogenesis. Strikingly, while resident TEs piRNAs seem to engage in a typical ping-pong amplification mechanism, sense and antisense HTT piRNAs instead overlap by ∼20 nt or do not display ping-pong signatures.
Our data shed light on the biogenesis and functions of the piRNAs in Rhodnius prolixus and reveal that piRNAs, but not the siRNA pathway, responded to HTTs that were recently transferred from vertebrate tetrapods to a hematophagous insect of medical relevance.
Piwi蛋白及相关的Piwi相互作用RNA(piRNA)共同构成一个监测系统,可保护动物基因组免受转座元件(TE)移动引起的DNA损伤。虽然该途径在果蝇中已有详细描述,但在更基础的昆虫中了解较少。罗得西亚锥蝽是一种半翅目昆虫,也是恰加斯病的主要传播媒介之一。它通过吸食蝙蝠、负鼠和松鼠猴获取了特定类别的水平转移转座子(HTT),从而为研究该物种中针对HTT的piRNA碱基反应提供了机会。
对映射到HTT和常驻转座元件的小RNA测序读数进行定量,并检查其是否具有piRNA特征,如1U和10A偏好、乒乓和相位特征。使用唯一映射的piRNA来鉴定罗得西亚锥蝽基因组中的piRNA簇。RNA测序数据用于定量转座子和Rp - PIWI基因的表达水平,并通过qRT - PCR进行验证。
通过分析罗得西亚锥蝽发育关键阶段piRNA簇表达和piRNA产生的时间动态,我们发现约28 nt长的piRNA峰值水平主要在胚胎发生期间与HTT和常驻TE表达的降低相关。引人注目的是,虽然常驻TE的piRNA似乎参与典型的乒乓扩增机制,但有义链和反义链HTT的piRNA反而重叠约20 nt或不显示乒乓特征。
我们的数据揭示了罗得西亚锥蝽中piRNA的生物发生和功能,并表明piRNA而非小干扰RNA(siRNA)途径对最近从脊椎动物四足动物转移到具有医学相关性的吸血昆虫的HTT有反应。