Identification of cyst wall proteins of the hypotrich ciliate Euplotes encysticus using a proteomics approach

Wang, Bangzheng; Niu, Tao; Bhatti, Muhammad Zeeshan; Chen, Fen-Fen; Wu, Lin; Chen, Jiwu

doi:10.1007/s12275-017-6422-7

Cited by 12 publications

(9 citation statements)

References 24 publications

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“…Ubiquitin-mediated proteolysis pathway analysis. During transformation of the trophonts into cysts, the cell morphology changes sharply accompanied by the rapid modifications in intracellular proteins 13,14 . Therefore, dramatic changes occurring in the proteins suggested an activation of the ubiquitin-proteasome system to meet the needs of protein degradation during cell morphological changes 8 .…”

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confidence: 99%

Novel insights into molecular mechanisms of Pseudourostyla cristata encystment using comparative transcriptomics

Pan

Niu

Bhatti

et al. 2019

Sci Rep

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The encystment of many ciliates is an advanced survival strategy against adversity and the most important reason for ciliates existence worldwide. However, the molecular mechanism for the encystment of free-living ciliates is poorly understood. Here, we performed comparative transcriptomic analysis of dormant cysts and trophonts from Pseudourostyla cristata using transcriptomics, qRT-PCR and bioinformatic techniques. We identified 2565 differentially expressed unigenes between the dormant cysts and the trophonts. The total number of differentially expressed genes in GO database was 1752. The differential unigenes noted to the GO terms were 1993. These differential categories were mainly related to polyamine transport, pectin decomposition, cytoplasmic translation, ribosome, respiratory chain, ribosome structure, ion channel activity, and RNA ligation. A total of 224 different pathways were mapped. Among them, 184 pathways were upregulated, while 162 were downregulated. Further investigation showed that the calcium and AMPK signaling pathway had important induction effects on the encystment. In addition, FOXO and ubiquitin-mediated proteolysis signaling pathway jointly regulated the encystment. Based on these findings, we propose a hypothetical signaling network that regulates Pseudourostyla cristata encystment. Overall, these results provide deeper insights into the molecular mechanisms of ciliates encystment and adaptation to adverse environments.of Strepkiella histriomuscorum has been studied in the encystment and trophont stages through biochip technology, which suggested that cyst formation is associated with Ribosomal L7, Ribosomal acidic P2, Cathepsin B, Cathepsin H, Ubiquitin, Ca 2+ -ATPase and Actin1 7 . Other studies investigated the differential proteins of the cysts and the trophonts from Pseudourostyla cristata (P. cristata) using shotgun LC-MS/MS, finding the association of fibrillarin-like rRNA methylase, methylmalonyl-coenzyme mutase, ADP ribosylation factor, Rab12, MAPK-related kinase and KR multi-domain proteins with cyst formation 8 . However, the reports of systematically studying the genes and signaling pathways involved in the formation of ciliate cysts at the molecular level are rare 8 . In this study, we have investigated the molecular mechanism underlying the cyst formation in ciliates using comparative transcriptomic analysis, quantitative real-time PCR (qRT-PCR) and bioinformatic techniques. Furthermore, we have focused on the analysis of several important signaling pathways related to the encystment. Therefore, we have proposed a hypothetical network that regulates Pseudourostyla cristata encystment. Our study generates novel insights into the molecular mechanisms of the cyst formation in free-living ciliates.www.nature.com/scientificreports www.nature.com/scientificreports/ dormant cysts of P. cristata (Fig. 4). Validation of results by transcriptome sequencing showed that mRNA outcomes were consistent with the transcriptomic data of trophonts and dormant cysts in P. cristata (Fig. 4).

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confidence: 99%

Novel insights into molecular mechanisms of Pseudourostyla cristata encystment using comparative transcriptomics

Pan

Niu

Bhatti

et al. 2019

Sci Rep

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“…Numerous reports have focused on the morphology of cysts 6 , 8 . Previously, we reported cyst wall proteins function and molecular mechanism of Euplotes encysticus under stress condition 9 . In this article, the life signal curve of the vegetative cells and the resting cysts from Euplotes encysticus was obtained by using the energy metabolism analysis.…”

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confidence: 96%

“…Fluorescent labeling was performed according to our previous method 9 with slight modification. In brief, 40 µL of the vegetative cells were added on a clean glass slide and excess water was removed, respectively.…”

Section: Methodsmentioning

confidence: 99%

New contribution to the morphology and molecular mechanism of Euplotes encysticus encystment

Chen

Xue

Pan

et al. 2018

Sci Rep

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Ciliated protists are a large group of single-cell eukaryotes, leading to the resting cysts in unfavorable environmental condition. However, the underlying molecular mechanism of encystment in the free-living ciliates is poorly understood. Here we show that the resting cysts are better than the vegetative cells of Euplotes encysticus in adverse survivor with respect to energy metabolism. Therefore scale identification of encystment-related proteins in Euplotes encysticus was investigated by iTRAQ analysis. We analyzed a total of 130 proteins, in which 19 proteins involving 12 upregulated and 7 downregulated proteins were associated with encystment in the resting cysts in comparison with the vegetative cells. Moreover, direct fluorescent labeling analysis showed that the vegetative cells treated with shRNA-β-tubulin recombinant E. coli accumulated a large number of granular materials, and dramatic cell morphology changes. Importantly, the cell membrane rupture phenomenon was observed after three weeks of shRNA-β-tubulin interference as compared to the control group. These results revealed that different proteins might play an important role in the process of the vegetative cells into the resting cysts. These results will help to reveal the morphological changes and molecular mechanism of resting cyst formation of ciliates.

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“…For the legend: macronuclei are not drawn in all stages for certain species; in Tetrahymena rostrata , the old macronucleus is degraded, while the new macronuclei are produced by the micronuclei; curved arrows indicate rotation and degree; mucocyst is secreted to form the cyst wall; degrading or regenerating ciliature is drawn as broken lines; vacuoles in Parentocirrus hortualis empty into the so-called contractile vacuole; pockets are disk-like structures consisting of unknown material. Tetrahymena rostrata ( McArdle et al, 1980 ; Segade et al, 2016 ); Holophrya teres ( Benčat’ová et al, 2020 ); Bursaria truncatella ( Beers, 1948 ); Rigidohymena quadrinucleata ( Benčat’ová and Tirjaková, 2017 ); Pleurotricha lanceolata ( Jeffries, 1956 ); Sterkiella histriomuscorum ( Adl and Berger, 1997 ); Stylonychia bifaria ( Verni and Rosati, 2011 ); Stylonychia mytilus ( Zhang and Pang, 1981 ); Halteria grandinella ( Foissner et al, 2007 ); Meseres corlissi ( Foissner et al, 2006 ; Müller, 2007 ); Parentocirrus hortualis ( Benčat’ová et al, 2016 ); Euplotes encysticus ( Rawlinson and Gates, 1985 ; Gu and Zhang, 1992 ; Wang B. et al, 2017 ); Phacodinium metchnikoffi ( Benčat’ová and Tirjaková, 2018 ); Blepharisma stoltei ( Repak, 1968 ).…”

Section: Structure Of Cystsmentioning

confidence: 99%

How Ciliated Protists Survive by Cysts: Some Key Points During Encystment and Excystment

Wang

Zhang

et al. 2022

Front. Microbiol.

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Forming cysts is a common and important bionomic strategy for microorganisms to persist in harsh environments. In ciliated protists, many species have been reported to form cysts when facing unfavorable conditions. Despite traditional studies on the morphological features of cysts and the chemical composition of cyst wall, recent research has focused more on the molecular mechanisms of encystment. The present work reviews studies on developmental features and molecular information of resting cysts in ciliates, and pays more attention to the following questions: what are the inducing factors of encystment and excystment? How does the cell change morphologically during these dynamic processes? And what molecular mechanisms underlie those changes? We also present and summarize the characteristics of cysts from diverse ciliate lineages in a phylogenetic framework, aiming to provide new perspectives for studies on adaptive evolution of unicellular eukaryotes.

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Identification of cyst wall proteins of the hypotrich ciliate Euplotes encysticus using a proteomics approach

Cited by 12 publications

References 24 publications

Novel insights into molecular mechanisms of Pseudourostyla cristata encystment using comparative transcriptomics

Novel insights into molecular mechanisms of Pseudourostyla cristata encystment using comparative transcriptomics

New contribution to the morphology and molecular mechanism of Euplotes encysticus encystment

How Ciliated Protists Survive by Cysts: Some Key Points During Encystment and Excystment

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