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Transposon targeting non-coding RNA transcription targets G/C-rich tracts and is facilitated by an intrinsically disordered protein in Tetrahymena
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Abstract
Tetrahymena is a well-established ciliate model organism, known for its nuclear dualism and unique mechanism for transposon defense, wherein transposons are kept within the highly heterochromatinized micronucleus (MIC), which is transcriptionally inert except during meiosis. The discovery of preferential transcription in the meiotic MIC at regions enriched with transposons and the identification of MIC transcription regulatory proteins emphasize the necessity to elucidate the underlying mechanisms driving this process. In this study, we demonstrate that G/C-rich tracts are overrepresented in meiotic non-coding RNA (ncRNA) transcription regions and that RNA polymerase II (Pol II) is highly enriched at and around these tracts. Further analysis revealed that Pol II's association with G/C-rich tracts is not abolished in cells lacking Rib1, a Mediator complex-associated protein essential for MIC ncRNA biogenesis. Nevertheless, in the absence of Rib1, Pol II showed abnormal association with the meiotic MIC chromatin, suggesting that Rib1 is critical for maintaining Pol II's binding specificity. Through Rib1 truncation analysis, we found that the intrinsically disordered region, which contains putative phase-separating peptides, is crucial for its function. Disruption of phase-separation, either by deleting these peptides or by treating cells with a phase-separation disruption reagent, leads to aberrant localization of both Rib1 and Pol II on the MIC chromatin, implicating that Rib1 likely facilitates the MIC ncRNA transcription via phase-separation.
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Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
Varadi, M., Bertoni, D., Magana, P., Paramval, U., Pidruchna, I., Radhakrishnan, M., Tsenkov, M., Nair, S., Mirdita, M., Yeo, J., et al. (2024) · Nucleic Acids Research · doi:10.1093/nar/gkad1011 · Cited by 2,052 · cited 2×
“Structural information for the Rib1 protein was predicted using AlphaFold and retrieved from UniProt (UniProt Consortium, 2025; Varadi et al., 2024).”
The Structure and Function of DNA G-Quadruplexes
Spiegel, J., Adhikari, S., & Balasubramanian, S. (2020) · Trends in Chemistry · doi:10.1016/j.trechm.2019.07.002 · Cited by 920 · cited 1×
“Given that G/C-rich DNA is prone to form G-quadruplex, a four-stranded secondary DNA structure (Lombardi and Londoño-Vallejo, 2020; Spiegel et al., 2020), and ample recent evidence has shown that G-quadruplex promotes transcription (Esain-Garcia et al., 2024; Wang et al., 2024), we then identified…”
A Field Guide to Eukaryotic Transposable Elements
Wells, J. N., & Feschotte, C. (2020) · Annual Review of Genetics · doi:10.1146/annurev-genet-040620-022145 · Cited by 686
A heterochromatin-dependent transcription machinery drives piRNA expression
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“Second, G-quadruplexes serve as binding platforms for transcription factors, which in turn facilitate transcription (Lago et al., 2021; Li et al., 2021b; Spiegel et al., 2021; Wulfridge et al., 2023; Yuan et al., 2023).”
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Metadata
- Authors
- Cai, X., Liu, Y., Li, J., Zhao, Z., Yao, X., Zhai, Z., Liucong, M., Liu, Y., Wang, X., Mochizuki, K., & Tian, M.
- Affiliations
- Ocean University of China (CN)
- Centre National de la Recherche Scientifique (FR)
- Corresponding author
- Tian, Miao
- Venue
- Science China Life Sciences
- Published
- 2026-02-26
- Volume
- 69
- Issue
- 7
- Pages
- 2156-2172
- ISSN
- 1674-7305 (ISSN-L)
- 1869-1889
- OpenAlex
- W7133187627
- Cited by
- 1
- References
- 6
- Citations by year
- 2026: 1
- FWCI
- 2.09
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Science China Life Sciences (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- Funders
- European Commission (HORIZON2020, 101024333)
- National Natural Science Foundation of China (32370451)
- Natural Science Foundation of Shandong Province (ZR2022JQ13)
- Fundamental Research Funds for the Central Universities (202241003)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Chromosomal and Genetic Variations
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Keywords
- Tetrahymena, micronuclei, transposon defense, RNA polymerase II, Mediator complex, intrinsically disordered proteins, phase separation, meiosis, heterochromatin, transcriptional regulation
- MeSH terms
- DNA Transposable Elements, Heterochromatin, Meiosis, RNA Polymerase II, Tetrahymena, Transcription, Genetic, Protozoan Proteins, RNA, Untranslated
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