"Evidence that Moderate Eviction of Spt5 and Promotion of Error-Free Tr" by Kathiresan Selvam, Baojin Ding et al.

Faculty Publications

Title

Evidence that Moderate Eviction of Spt5 and Promotion of Error-Free Transcriptional Bypass by Rad26 Facilitates Transcription Coupled Nucleotide Excision Repair

Authors

Kathiresan Selvam, Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA.
Baojin Ding, Department of Biology, University of Louisiana at Lafayette, 410 E. St. Mary Blvd, Lafayette, LA 70503, USA.
Rahul Sharma, National Hansen's Disease Program, Laboratory Research Branch at Louisiana State University, 3519E School of Veterinary Medicine, Baton Rouge, LA 70803, USA.
Shisheng Li, Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA. Electronic address: shli@lsu.edu.

Document Type

Article

Publication Date

3-29-2019

Abstract

Transcription coupled repair (TC-NER) is a subpathway of nucleotide excision repair triggered by stalling of RNA polymerase at DNA lesions. It has been suspected that transcriptional misincorporations of certain nucleotides opposite lesions that result in irreversible transcription stalling might be important for TC-NER. However, the spectra of nucleotide misincorporations opposite UV photoproducts and how they are implicated in transcriptional stalling and TC-NER in the cell remain unknown. Rad26, a low abundant yeast protein, and its human homolog CSB have been proposed to facilitate TC-NER in part by positioning and stabilizing stalling of RNA polymerase II (RNAPII) at DNA lesions. Here, we found that substantial AMPs but no other nucleotides are transcriptionally misincoporated and extended opposite UV photoproducts and adjacent bases in Saccharomyces cerevisiae. Rad26 does not significantly affect either the misincorporation or extension of AMPs. At normally low or moderately increased levels, Rad26 promotes error-free transcriptional bypass and TC-NER of UV photoproducts. However, Rad26 completely loses these functions when it is overexpressed to ~1/3 the level of RNAPII molecules. Also, Rad26 does not directly displace RNAPII but constitutively evicts Spt5, a key transcription elongation factor and TC-NER repressor, from the chromatin. Our results indicate that transcriptional nucleotide misincorporation is not implicated in TC-NER, and moderate eviction of Spt5 and promotion of error-free transcriptional bypass of DNA lesions by Rad26 facilitates TC-NER.

Publication Source (Journal or Book title)

Journal of molecular biology

First Page

1322

Last Page

1338

Recommended Citation

Selvam, K., Ding, B., Sharma, R., & Li, S. (2019). Evidence that Moderate Eviction of Spt5 and Promotion of Error-Free Transcriptional Bypass by Rad26 Facilitates Transcription Coupled Nucleotide Excision Repair. Journal of molecular biology, 431 (7), 1322-1338. https://doi.org/10.1016/j.jmb.2019.02.010

This document is currently not available here.

COinS