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RNA聚合酶II和III募集至snRNA启动子的不同机制的结构见解

Structural insights into distinct mechanisms of RNA polymerase II and III recruitment to snRNA promoters.

作者信息

Shah Syed Zawar, Perry Thomas N, Graziadei Andrea, Cecatiello Valentina, Kaliyappan Thangavelu, Misiaszek Agata D, Müller Christoph W, Ramsay Ewan P, Vannini Alessandro

机构信息

Human Technopole, Milan, Italy.

Institute of Cancer Research, London, UK.

出版信息

Nat Commun. 2025 Jan 2;16(1):141. doi: 10.1038/s41467-024-55553-8.

DOI:10.1038/s41467-024-55553-8
PMID:39747245
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11696126/
Abstract

RNA polymerase III (Pol III) transcribes short, essential RNAs, including the U6 small nuclear RNA (snRNA). At U6 snRNA genes, Pol III is recruited by the snRNA Activating Protein Complex (SNAPc) and a Brf2-containing TFIIIB complex, forming a pre-initiation complex (PIC). Uniquely, SNAPc also recruits Pol II at the remaining splicesosomal snRNA genes (U1, 2, 4 and 5). The mechanism of SNAPc cross-polymerase engagement and the role of the SNAPC2 and SNAPC5 subunits remain poorly defined. Here, we present cryo-EM structures of the full-length SNAPc-containing Pol III PIC assembled on the U6 snRNA promoter in the open and melting states at 3.2-4.2 Å resolution. The structural comparison revealed differences with the Saccharomyces cerevisiae Pol III PIC and the basis of selective SNAPc engagement within Pol III and Pol II PICs. Additionally, crosslinking mass spectrometry localizes SNAPC2 and SNAPC5 near the promoter DNA, expanding upon existing descriptions of snRNA Pol III PIC structure.

摘要

RNA聚合酶III（Pol III）转录短的必需RNA，包括U6小核RNA（snRNA）。在U6 snRNA基因处，Pol III由snRNA激活蛋白复合物（SNAPc）和含Brf2的TFIIIB复合物招募，形成预起始复合物（PIC）。独特的是，SNAPc还在其余剪接体snRNA基因（U1、2、4和5）处招募Pol II。SNAPc跨聚合酶结合的机制以及SNAPC2和SNAPC5亚基的作用仍不清楚。在这里，我们展示了在U6 snRNA启动子上组装的全长含SNAPc的Pol III PIC在开放和解链状态下的冷冻电镜结构，分辨率为3.2 - 4.2埃。结构比较揭示了与酿酒酵母Pol III PIC的差异以及SNAPc在Pol III和Pol II PIC中选择性结合的基础。此外，交联质谱法将SNAPC2和SNAPC5定位在启动子DNA附近，扩展了对snRNA Pol III PIC结构的现有描述。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/4d694bbfad6b/41467_2024_55553_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/e0ceeb09e4b4/41467_2024_55553_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/8037c9bf1486/41467_2024_55553_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/d20c84aa5a2a/41467_2024_55553_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/af8b71220418/41467_2024_55553_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/26c4a12d643e/41467_2024_55553_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/4d694bbfad6b/41467_2024_55553_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/e0ceeb09e4b4/41467_2024_55553_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/8037c9bf1486/41467_2024_55553_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/d20c84aa5a2a/41467_2024_55553_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/af8b71220418/41467_2024_55553_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/26c4a12d643e/41467_2024_55553_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d32e/11696126/4d694bbfad6b/41467_2024_55553_Fig6_HTML.jpg

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