Proteomic identification and expression of oral apparatus constituents in cell regeneration of giant ciliate Stentor coeruleus (strain WHEL)

Wei, Wei; Jiang, Chuanqi; Yang, Wentao; Miao, Wei; Xiong, Jie

doi:10.1016/j.gene.2020.144624

Cited by 8 publications

(33 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…S. coeruleus (strain WHEL) [5] cells were grown at 20 • C in the dark in Modified Stentor Medium (MSM) [11], supplemented with Chlorogonium elongatum as living prey. Wild-type (WT) and the transformed strains of Synechocystis 6803 were grown at 30 • C in liquid or solid Blue-Green-11 (BG-11) medium at a light intensity of 40 µmoL photons m −2 s −1 in ambient air.…”

Section: Cell Culture and Growth Conditionsmentioning

confidence: 99%

“…To clone Stentor gene sequences, homologs were identified by best-reciprocal BLAST between the S. coeruleus WHEL [5] transcriptome and the S. coeruleus WM001 genome [25]. Target gene sequences were obtained by PCR amplification from S. coeruleus WHEL genomic DNA.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…It consists of thousands of long cilia organized into membranelles that compose the membranellar band (MB) [2]. In S. coeruleus, OA shedding can be induced by short urea treatment [3]; after removal and washing, the cells can completely regenerate the OA and finally restore the feeding ability within 9 h, progressing through a series of well-characterized morphological steps [1,4,5]. Single-cell transcriptome analysis revealed that the morphological steps in OA regeneration are driven by a complex regulatory system that includes centriole assembly, ciliogenesis, signaling, cell cycle regulation, transcription, and RNA binding in S. coeruleus [4].…”

Section: Introductionmentioning

confidence: 99%

“…Single-cell transcriptome analysis revealed that the morphological steps in OA regeneration are driven by a complex regulatory system that includes centriole assembly, ciliogenesis, signaling, cell cycle regulation, transcription, and RNA binding in S. coeruleus [4]. Using proteomic identification, we identified 882 OA-associated proteins in extracts from the Microorganisms 2021, 9, 176 2 of 12 shedding OA of S. coeruleus; most were highly and continuously expressed in OA regeneration [5]. However, few studies so far have focused on the function of genes involved in OA regeneration.…”

Section: Introductionmentioning

confidence: 99%

“…We established a novel RNAi method for the constitutive expression of exogenous gene dsRNA in Synechocystis 6803 and successfully applied this method to knock down a previous identified S. coeruleus-specific gene (named SCSG1), known to be associated with OA regeneration [5], and verified its function in ciliogenesis of OA regeneration.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

RNA Interference by Cyanobacterial Feeding Demonstrates the SCSG1 Gene Is Essential for Ciliogenesis during Oral Apparatus Regeneration in Stentor

et al. 2021

Self Cite

View full text Add to dashboard Cite

In the giant ciliate Stentor coeruleus, oral apparatus (OA) regeneration is an experimentally tractable regeneration paradigm that occurs via a series of morphological steps. OA regeneration is thought to be driven by a complex regulatory system that orchestrates the temporal expression of conserved and specific genes. We previously identified a S. coeruleus-specific gene (named SCSG1) that was significantly upregulated during the ciliogenesis stages of OA regeneration, with an expression peak at the stage of the first OA cilia appearance. We established a novel RNA interference (RNAi) method through cyanobacteria Synechocystis sp. PCC6803 feeding in S. coeruleus. The expression of SCSG1 gene was significantly knocked down by using this method and induced abnormal ciliogenesis of OA regeneration in S. coeruleus, suggesting that SCSG1 is essential for OA regeneration in S. coeruleus. This novel RNAi method by cyanobacterial feeding has potential utility for studying other ciliates.

show abstract

Section: Cell Culture and Growth Conditionsmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

RNA Interference by Cyanobacterial Feeding Demonstrates the SCSG1 Gene Is Essential for Ciliogenesis during Oral Apparatus Regeneration in Stentor

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

Determining protein polarization proteome-wide using physical dissection of individual Stentor coeruleus cells

et al. 2022

View full text Add to dashboard Cite

Modular, Cascade-like Transcriptional Program of Regeneration inStentor

Sood

Lin

McGillivary

et al. 2021

Preprint

View full text Add to dashboard Cite

The giant ciliate Stentor coeruleus is a classical model system for studying regeneration and morphogenesis at the level of a single cell. The anterior of the cell is marked by an array of cilia, known as the oral apparatus, which can be induced to shed and regenerate in a series of reproducible morphological steps, previously shown to require transcription. If a cell is cut in half, each half will regenerate an intact cell, including a new oral apparatus in the posterior half. We used RNAseq to assay the dynamic changes in Stentors transcriptome during regeneration, after both oral apparatus shedding and bisection, allowing us to identify distinct temporal waves of gene expression including kinases, RNA binding proteins, centriole biogenesis factors, and orthologs of human ciliopathy genes implicated in Meckel and Joubert syndromes. By comparing transcriptional profiles of different regeneration events in the same species, we were able to identify distinct modules of gene expression corresponding to oral apparatus regeneration, posterior holdfast regeneration, and recovery after wounding. By measuring gene expression in cells in which translation is blocked, we show that the sequential waves of gene expression involve a cascade mechanism in which later waves of expression are triggered by translation products of early-expressed genes. Among the early-expressed genes, we identified an E2F transcription factor and the conserved RNA binding protein Pumilio as potential regulators of regeneration based on the expression pattern of their predicted target genes. This work allows us to classify regeneration genes into groups based on their potential role for regeneration in distinct cell regeneration paradigms, and provides new insight into how a single cell can coordinate complex morphogenetic pathways to regenerate missing structures.

show abstract

Proteomic identification and expression of oral apparatus constituents in cell regeneration of giant ciliate Stentor coeruleus (strain WHEL)

Cited by 8 publications

References 43 publications

RNA Interference by Cyanobacterial Feeding Demonstrates the SCSG1 Gene Is Essential for Ciliogenesis during Oral Apparatus Regeneration in Stentor

RNA Interference by Cyanobacterial Feeding Demonstrates the SCSG1 Gene Is Essential for Ciliogenesis during Oral Apparatus Regeneration in Stentor

Determining protein polarization proteome-wide using physical dissection of individual Stentor coeruleus cells

Modular, Cascade-like Transcriptional Program of Regeneration inStentor

Contact Info

Product

Resources

About