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Journals / Turkish Journal of Biology / 2021 / Cilt: 45 - Sayı: 3

Reconstitution of Pol II (G) responsive form of the human Mediator complex

Murat Alper CEVHER
Pages
253–261
Publication Date
2022-10-17
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Our results show complete recovery of Pol II (G) dependent transcription both with full 30-subunit Mediator and also with 15-subunit recombinant Mediator that we synthesized.

📌 3 Key Takeaways

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Objective & Hypothesis:RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression.
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Key Findings & Data:Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro.
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Conclusion & Impact:Our results show complete recovery of Pol II (G) dependent transcription both with full 30-subunit Mediator and also with 15-subunit recombinant Mediator that we synthesized.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression. Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro. Here we report for the first time, the identification and the generation of a 15-subunit human Mediator complex via the novel multibac baculovirus expression system that is fully responsive to Pol II (G). Our results show complete recovery of Pol II (G) dependent transcription both with full 30-subunit Mediator and also with 15-subunit recombinant Mediator that we synthesized. Moreover, we also show that the recombinant Mediator interacts with Pol II (G) as well. These results enlighten us towards understanding how a certain population of Pol II that is involved in selected gene regulation is activated by Mediator complex.

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Cite This Article

CEVHER, M. A. (2022). Reconstitution of Pol II (G) responsive form of the human Mediator complex. Turkish Journal of Biology, Cilt: 45 - Sayı: 3, 253–261. https://ajindex.com/articles/583240

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References (4)

  1. Allen BL, Taatjes DJ (2015). The Mediator complex: a central integrator of transcription. Nature Reviews Molecular Cell Biology 16 (3): 155-166. doi: 10.1038/nrm3951 Blazek E, Mittler G, Meisterernst M (2005). The mediator of RNA polymerase II. Chromosoma 113 (8): 399-408. doi: 10.1007/ s00412-005-0329-5
  2. Cevher MA, Shi Y, Li D, Chait BT, Malik S et al. (2014). Reconstitution of active human core Mediator complex reveals a critical role of the MED14 subunit. Nature Structural and Molecular Biology 21 (12): 1028-1034. doi: 10.1038/nsmb.2914 Cheng B, Li T, Rahl PB, Adamson TE, Loudas NB et al. (2012). Functional association of gdown1 with RNA polymerase II poised on human genes. Molecular Cell 45 (1): 38-50. doi: 10.1016/j.molcel.2011.10.022 FagerbergL, Hallstrom BM, Oksvold P, Kampf C, Djureinovic D et al. (2014). Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibodybased proteomics. Molecular and Cellular Proteomics 13 (2): 397-406. doi: 10.1074/mcp.M113.035600 Guo J, Turek ME, Price DH (2014). Regulation of RNA polymerase II termination by phosphorylation of Gdown1. The Journal of Biological Chemistry 289 (18): 12657-12665. doi: 10.1074/jbc. M113.537662 Hu X, Malik S, Negroiu CC, Hubbard K, Velalar CN et al. (2006). A Mediator-responsive form of metozoan RNA polymerase II. Proceedings of the National Academy of Sciences of the United States of America 103 (25): 9506-9511. doi: 10.1073/ pnas.0603702103 Jeronimo C, Robert F (2017). The Mediator complex: at the nexus of RNA polymerase II transcription. Trends in Cell Biology 27 (10): 765-783. doi: 10.1016/j.tcb.2017.07.001 Jishage M, Malik S, Wagner U, Uberheide B, Ishihama Y et al. (2012). Transcriptional regulation by pol II(G) involving mediator and competitive interactions of gdown1 and tfiif with pol II. Molecular Cell 45 (1): 51-63. doi: 10.1016/j.molcel.2011.12.014 Jishage M, Yu X, Shi Y, Ganesan SJ, Chen WY et al. (2018). Architecture of Pol II(G) and molecular mechanism of transcription regulation by Gdown1. Nature Structural and Molecular Biology 25 (9): 859-867. doi: 10.1038/s41594-018- 0118-5
  3. Kim YJ, Björklund S, Li Y, Sayre MH, Kornberg RD (1994). A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA polymerase II. Cell 77 (4): 599-608. doi: 10.1016/0092- 8674(94)90221-6
  4. Lacombe T, PohSL, Gine Barbey R, Kuras L (2013). Mediator is an intrinsic component of the basal RNA polymerase II