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Article: Intrinsic cleavage of RNA polymerase II adopts a nucleobase-independent mechanism assisted by transcript phosphate

TitleIntrinsic cleavage of RNA polymerase II adopts a nucleobase-independent mechanism assisted by transcript phosphate
Authors
Issue Date2019
PublisherNature Publishing Group. The Journal's web site is located at https://www.nature.com/natcatal/
Citation
Nature Catalysis, 2019, v. 2 n. 3, p. 228-235 How to Cite?
AbstractRNA polymerase II (Pol II) utilizes the same active site for polymerization and intrinsic cleavage. Pol II proofreads the nascent transcript via its intrinsic nuclease activity to maintain high transcriptional fidelity critical for cell growth and viability. The detailed catalytic mechanism of intrinsic cleavage remains unknown. Here, we combined ab initio quantum mechanics/molecular mechanics studies and biochemical cleavage assays to show that Pol II utilizes downstream phosphate oxygen to activate the attacking nucleophile in hydrolysis, while the newly formed 3′-end is protonated through active-site water without a defined general acid. Experimentally, alteration of downstream phosphate oxygen either by 2′-5′ sugar linkage or stereo-specific thio-substitution of phosphate oxygen drastically reduced cleavage rate. We showed by N7-modification that guanine nucleobase is not directly involved as an acid–base catalyst. Our proposed mechanism provides important insights into the intrinsic transcriptional cleavage reaction, an essential step in transcriptional fidelity control.
Persistent Identifierhttp://hdl.handle.net/10722/272138
ISSN
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorTse, CKM-
dc.contributor.authorXu, J-
dc.contributor.authorXu, L-
dc.contributor.authorSheong, FK-
dc.contributor.authorWang, S-
dc.contributor.authorChow, HY-
dc.contributor.authorGao, X-
dc.contributor.authorLi, X-
dc.contributor.authorCheung, PPH-
dc.contributor.authorWang, D-
dc.contributor.authorZhang, Y-
dc.contributor.authorHuang, X-
dc.date.accessioned2019-07-20T10:36:24Z-
dc.date.available2019-07-20T10:36:24Z-
dc.date.issued2019-
dc.identifier.citationNature Catalysis, 2019, v. 2 n. 3, p. 228-235-
dc.identifier.issn2520-1158-
dc.identifier.urihttp://hdl.handle.net/10722/272138-
dc.description.abstractRNA polymerase II (Pol II) utilizes the same active site for polymerization and intrinsic cleavage. Pol II proofreads the nascent transcript via its intrinsic nuclease activity to maintain high transcriptional fidelity critical for cell growth and viability. The detailed catalytic mechanism of intrinsic cleavage remains unknown. Here, we combined ab initio quantum mechanics/molecular mechanics studies and biochemical cleavage assays to show that Pol II utilizes downstream phosphate oxygen to activate the attacking nucleophile in hydrolysis, while the newly formed 3′-end is protonated through active-site water without a defined general acid. Experimentally, alteration of downstream phosphate oxygen either by 2′-5′ sugar linkage or stereo-specific thio-substitution of phosphate oxygen drastically reduced cleavage rate. We showed by N7-modification that guanine nucleobase is not directly involved as an acid–base catalyst. Our proposed mechanism provides important insights into the intrinsic transcriptional cleavage reaction, an essential step in transcriptional fidelity control.-
dc.languageeng-
dc.publisherNature Publishing Group. The Journal's web site is located at https://www.nature.com/natcatal/-
dc.relation.ispartofNature Catalysis-
dc.titleIntrinsic cleavage of RNA polymerase II adopts a nucleobase-independent mechanism assisted by transcript phosphate-
dc.typeArticle-
dc.identifier.emailLi, X: xuechenl@hku.hk-
dc.identifier.authorityLi, X=rp00742-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1038/s41929-019-0227-5-
dc.identifier.pmid31179024-
dc.identifier.pmcidPMC6548511-
dc.identifier.scopuseid_2-s2.0-85061368443-
dc.identifier.hkuros299161-
dc.identifier.volume2-
dc.identifier.issue3-
dc.identifier.spage228-
dc.identifier.epage235-
dc.identifier.isiWOS:000461099900013-
dc.publisher.placeUnited Kingdom-

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