8TVS

Cryo-EM structure of backtracked Pol II in complex with Rad26

Summary for 8TVS

• Entry DOI: 10.2210/pdb8tvs/pdb
• EMDB information: 41650
• Descriptor: DNA-directed RNA polymerase subunit, DNA-directed RNA polymerases II subunit RPABC5, DNA-directed RNA polymerase II subunit RPB11, ... (18 entities in total)
• Functional Keywords: rna polymerase ii, rad26, cpd lesion, transcription-coupled dna repair, backtracked polymerase, transcription, transcription-dna-rna complex, transcription/dna/rna
• Biological source: Saccharomyces cerevisiae; More
• Total number of polymer chains: 16
• Total formula weight: 585952.14

Authors

Sarsam, R.D.,Lahiri, I.,Leschziner, A.E. (deposition date: 2023-08-18, release date: 2024-01-24, Last modification date: 2024-10-16)

Primary citation

Sarsam, R.D.,Xu, J.,Lahiri, I.,Gong, W.,Li, Q.,Oh, J.,Zhou, Z.,Hou, P.,Chong, J.,Hao, N.,Li, S.,Wang, D.,Leschziner, A.E.
Elf1 promotes Rad26's interaction with lesion-arrested Pol II for transcription-coupled repair.
Proc.Natl.Acad.Sci.USA, 121:e2314245121-e2314245121, 2024

PubMed Abstract: Transcription-coupled nucleotide excision repair (TC-NER) is a highly conserved DNA repair pathway that removes bulky lesions in the transcribed genome. Cockayne syndrome B protein (CSB), or its yeast ortholog Rad26, has been known for decades to play important roles in the lesion-recognition steps of TC-NER. Another conserved protein ELOF1, or its yeast ortholog Elf1, was recently identified as a core transcription-coupled repair factor. How Rad26 distinguishes between RNA polymerase II (Pol II) stalled at a DNA lesion or other obstacles and what role Elf1 plays in this process remains unknown. Here, we present cryo-EM structures of Pol II-Rad26 complexes stalled at different obstacles that show that Rad26 uses a common mechanism to recognize a stalled Pol II, with additional interactions when Pol II is arrested at a lesion. A cryo-EM structure of lesion-arrested Pol II-Rad26 bound to Elf1 revealed that Elf1 induces further interactions between Rad26 and a lesion-arrested Pol II. Biochemical and genetic data support the importance of the interplay between Elf1 and Rad26 in TC-NER initiation. Together, our results provide important mechanistic insights into how two conserved transcription-coupled repair factors, Rad26/CSB and Elf1/ELOF1, work together at the initial lesion recognition steps of transcription-coupled repair.

PubMed: 38194460
DOI: 10.1073/pnas.2314245121

Experimental method

ELECTRON MICROSCOPY (4.4 Å)