Identification and subcellular localization analysis of CCTα in microsporidian Nosema bombycis

Yao, Mingshuai; Wang, Runpeng; Chen, Ying; He, Ping; Wei, Erjun; Zhu, Feng; Wang, Qiang; Zhang, Shujun; Tang, Xudong; Shen, Zhongyuan

doi:10.1016/j.meegid.2022.105309

Cited by 3 publications

(4 citation statements)

References 29 publications

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“…During the proliferative phase, the cytoskeletal proteins such as actin and tubulin are actively synthesized to enable cell division, as well as the intracellular transport of the components of the spore wall that starts forming at this stage [ 33 ]. These lines of evidence are similar to our previous findings on NbCCTα [ 24 ] and indicate that NbCCTζ may have a function like NbCCTα in the folding of tubulin and actin.…”

Section: Discussionsupporting

confidence: 91%

“…Our previous work has proved that RNAi can downregulate the expression of NbCCTα [ 24 ]. In this study, after the knockdown of NbCCTα or NbCCTζ , the relative expression of NbCCTζ or NbCCTα was detected by RT-qPCR using NbssrRNA as a reference gene.…”

Section: Resultsmentioning

confidence: 99%

“…The transcriptional level of NbCCTα was downregulated after the knockdown of the NbCCTζ gene at 24 h, 72 h, and 96 h, whereas it was significantly upregulated at 48 h; moreover, the proliferation of N. bombycis was significantly inhibited, indicating that the NbCCTζplays a crucial role in the proliferation of N. bombycis . Our previous studies have revealed that NbCCTα is related to cell division, and NbCCTδ plays an efficient role in keeping the integrity of the cytoskeleton [ 24 , 37 ]. Combining the results of this study with those of our previous work, we summarize our knowledge of the expression pattern and subcellular localization of CCTζ, CCTδ, and CCTα in N. bombycis in Table S1 , which may help to perform further research on the function and relationship of CCT subunits in microsporidia.…”

Section: Discussionmentioning

confidence: 99%

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The Function of Different Subunits of the Molecular Chaperone CCT in the Microsporidium Nosema bombycis: NbCCTζ Interacts with NbCCTα

Xu,

Chen,

et al. 2024

JoF

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Chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone protein that consists of eight completely different subunits and assists in the folding of newly synthesized peptides. The zeta subunit of CCT is a regulatory factor for the folding and assembly of cytoskeletal proteins as individuals or complexes. In this study, the zeta subunit of Nosema bombycis (NbCCTζ) is identified for the first time. The complete ORF of the NbCCTζ gene is 1533 bp in length and encodes a 510 amino acid polypeptide. IFA results indicate that NbCCTζ is colocalized with actin and β-tubulin in the cytoplasm during the proliferative phase and that NbCCTζ is completely colocalized with NbCCTα in the cytoplasm of N. bombycis throughout the entire life cycle. Furthermore, the yeast two-hybrid assay revealed that the NbCCTζ interacts with NbCCTα. The transcriptional level of NbCCTζ is significantly downregulated by knocking down the NbCCTα gene, while the transcriptional level of NbCCTα is downregulated after knocking down the NbCCTζ gene. These results suggest that NbCCTζ may play a vital role in the proliferation of N. bombycis by coordinating with NbCCTα.

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Section: Discussionsupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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The Function of Different Subunits of the Molecular Chaperone CCT in the Microsporidium Nosema bombycis: NbCCTζ Interacts with NbCCTα

Xu,

Chen,

et al. 2024

JoF

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“…Most of these studies are focused on the search for parasite genes as effective targets for suppression the growth of N. bombycis using RNA interference (RNAi) methods [ 8 , 9 , 10 , 11 ]. Along with double-stranded RNA (dsRNA) and small RNA (siRNA) molecules used to specifically suppress the expression of microsporidia genes, metal (silver) nanoparticles may also be effective in the treatment of infections caused by N. bombycis [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Construction of scFv Antibodies against the Outer Loops of the Microsporidium Nosema bombycis ATP/ADP-Transporters and Selection of the Fragment Efficiently Inhibiting Parasite Growth

Dolgikh

Senderskiy

Timofeev

et al. 2022

IJMS

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Traditional sanitation practices remain the main strategy for controlling Bombyx mori infections caused by microsporidia Nosema bombycis. This actualizes the development of new approaches to increase the silkworm resistance to this parasite. Here, we constructed a mouse scFv library against the outer loops of N. bombycis ATP/ADP carriers and selected nine scFv fragments to the transporter, highly expressed in the early stages of the parasite intracellular growth. Expression of selected scFv genes in Sf9 cells, their infection with different ratios of microsporidia spores per insect cell, qPCR analysis of N. bombycis PTP2 and Spodoptera frugiperda COXI transcripts in 100 infected cultures made it possible to select the scFv fragment most effectively inhibiting the parasite growth. Western blot analysis of 42 infected cultures with Abs against the parasite β-tubulin confirmed its inhibitory efficiency. Since the VL part of this scFv fragment was identified as a human IgG domain retained from the pSEX81 phagemid during library construction, its VH sequence should be a key antigen-recognizing determinant. Along with the further selection of new recombinant Abs, this suggests the searching for its natural mouse VL domain or “camelization” of the VH fragment by introducing cysteine and hydrophilic residues, as well as the randomization of its CDRs.

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Glucose-regulated protein 78 regulates the subunit-folding of the CCT complex by modulating gene expression and protein interaction in the microsporidian Nosema bombycis

Xu,

Xiao,

et al. 2025

International Journal of Biological Macromolecules

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Identification and subcellular localization analysis of CCTα in microsporidian Nosema bombycis

Cited by 3 publications

References 29 publications

The Function of Different Subunits of the Molecular Chaperone CCT in the Microsporidium Nosema bombycis: NbCCTζ Interacts with NbCCTα

The Function of Different Subunits of the Molecular Chaperone CCT in the Microsporidium Nosema bombycis: NbCCTζ Interacts with NbCCTα

Construction of scFv Antibodies against the Outer Loops of the Microsporidium Nosema bombycis ATP/ADP-Transporters and Selection of the Fragment Efficiently Inhibiting Parasite Growth

Glucose-regulated protein 78 regulates the subunit-folding of the CCT complex by modulating gene expression and protein interaction in the microsporidian Nosema bombycis

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