束带栉水母的粘附促进了由 cAMP 信号驱动的发育命运的快速改变。

IF 3.7 2区 生物学 Q2 MICROBIOLOGY mSphere Pub Date : 2024-10-29 Epub Date: 2024-09-24 DOI:10.1128/msphere.00617-24
Shane Denecke, Madeline F Malfara, Kelly R Hodges, Nikki A Holmes, Andre R Williams, Julia H Gallagher-Teske, Julia M Pascarella, Abigail M Daniels, Geert Jan Sterk, Rob Leurs, Gordon Ruthel, Rachel Hoang, Megan L Povelones, Michael Povelones
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引用次数: 0

摘要

锥虫是导致人类和动物疾病的单细胞寄生虫。通常情况下,昆虫宿主的定殖是传播的必要条件。寄生虫通过单鞭毛稳定地附着在昆虫组织上,同时发生分化和形态变化。虽然附着是锥虫生命周期中的一个保守阶段,但其分子机制还不十分清楚。为了研究这一过程,我们详细阐述了一个体外模型,在该模型中,锥虫Crithidia fasciculata的游动形态在粘附到人工基底后迅速分化。对从游动型过渡到附着型细胞的实时成像显示,寄生虫经历了一系列确定的事件,包括鞭毛基部附近的初始附着,紧接着是鞭毛缩短、细胞变圆,以及在缩短的鞭毛顶端和基质之间形成类似半膜的附着斑。游动寄生虫与附着寄生虫的定量蛋白质组学研究表明,以环磷酸腺苷(cAMP)为基础的信号蛋白的调控存在差异。我们在游动的 C. fasciculata 的鞭毛上定位到了其中两种蛋白,但在附着细胞的缩短鞭毛上却发现这两种蛋白都不存在。药物抑制 cAMP 磷酸二酯酶可增加细胞中的 cAMP 水平并阻止附着。此外,用抑制剂处理不会影响游动细胞或已附着细胞的生长速度,这表明抑制剂的作用仅限于附着早期的一个关键窗口。这些数据表明,C. fasciculata 的附着需要 cAMP 信号传导,在分化和附着过程中,鞭毛信号传导结构域可能会发生重组。在昆虫体内,寄生虫附着在组织上,有时作为附着细胞分裂,或产生运动性、传染性形式。对附着的意义和细胞机制的研究相对较少。在这里,我们利用一种能牢固附着在人造表面上的锥虫模型来更好地了解这一过程。这种附着再现了在体内观察到的情况,可用于确定附着的阶段和形态特征以及影响附着效率的条件。我们发现了在游动或附着的寄生虫中都富集的蛋白质,支持环磷酸腺苷信号通路在从游动到附着的过渡中发挥作用。由于这一途径已经与锥虫的环境感知和发育过渡有关,我们的数据为寄生虫在昆虫宿主体内生存所需的活动提供了新的见解。
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Adhesion of Crithidia fasciculata promotes a rapid change in developmental fate driven by cAMP signaling.

Trypanosomatids are single-celled parasites responsible for human and animal disease. Typically, colonization of an insect host is required for transmission. Stable attachment of parasites to insect tissues via their single flagellum coincides with differentiation and morphological changes. Although attachment is a conserved stage in trypanosomatid life cycles, the molecular mechanisms are not well understood. To study this process, we elaborate upon an in vitro model in which the swimming form of the trypanosomatid Crithidia fasciculata rapidly differentiates following adhesion to artificial substrates. Live imaging of cells transitioning from swimming to attached shows parasites undergoing a defined sequence of events, including an initial adhesion near the base of the flagellum immediately followed by flagellar shortening, cell rounding, and the formation of a hemidesmosome-like attachment plaque between the tip of the shortened flagellum and the substrate. Quantitative proteomics of swimming versus attached parasites suggests differential regulation of cyclic adenosine monophosphate (cAMP)-based signaling proteins. We have localized two of these proteins to the flagellum of swimming C. fasciculata; however, both are absent from the shortened flagellum of attached cells. Pharmacological inhibition of cAMP phosphodiesterases increased cAMP levels in the cell and prevented attachment. Further, treatment with inhibitor did not affect the growth rate of either swimming or established attached cells, indicating that its effect is limited to a critical window during the early stages of adhesion. These data suggest that cAMP signaling is required for attachment of C. fasciculata and that flagellar signaling domains may be reorganized during differentiation and attachment.IMPORTANCETrypanosomatid parasites cause significant disease burden worldwide and require insect vectors for transmission. In the insect, parasites attach to tissues, sometimes dividing as attached cells or producing motile, infectious forms. The significance and cellular mechanisms of attachment are relatively unexplored. Here, we exploit a model trypanosomatid that attaches robustly to artificial surfaces to better understand this process. This attachment recapitulates that observed in vivo and can be used to define the stages and morphological features of attachment as well as conditions that impact attachment efficiency. We have identified proteins that are enriched in either swimming or attached parasites, supporting a role for the cyclic AMP signaling pathway in the transition from swimming to attached. As this pathway has already been implicated in environmental sensing and developmental transitions in trypanosomatids, our data provide new insights into activities required for parasite survival in their insect hosts.

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来源期刊
mSphere
mSphere Immunology and Microbiology-Microbiology
CiteScore
8.50
自引率
2.10%
发文量
192
审稿时长
11 weeks
期刊介绍: mSphere™ is a multi-disciplinary open-access journal that will focus on rapid publication of fundamental contributions to our understanding of microbiology. Its scope will reflect the immense range of fields within the microbial sciences, creating new opportunities for researchers to share findings that are transforming our understanding of human health and disease, ecosystems, neuroscience, agriculture, energy production, climate change, evolution, biogeochemical cycling, and food and drug production. Submissions will be encouraged of all high-quality work that makes fundamental contributions to our understanding of microbiology. mSphere™ will provide streamlined decisions, while carrying on ASM''s tradition for rigorous peer review.
期刊最新文献
Erratum for Choi et al., "Human saliva modifies growth, biofilm architecture, and competitive behaviors of oral streptococci". mSphere of Influence: Venturing outside the biology canon with sex and gender. Pneumococcal extracellular vesicles mediate horizontal gene transfer via the transformation machinery. Adaptation to zinc restriction in Streptococcus agalactiae: role of the ribosomal protein and zinc-importers regulated by AdcR. Analytical measuring interval, linearity, and precision of serology assays for detection of SARS-CoV-2 antibodies according to CLSI guidelines.
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