title: Cockayne syndrome B protein regulates recruitment of the Elongin A ubiquitin ligase to sites of DNA damage.
abstract: Elongin A performs dual functions as the transcriptionally active subunit of RNA polymerase II (Pol II) elongation factor Elongin and as the substrate recognition subunit of a Cullin-RING E3 ubiquitin ligase that ubiquitylates Pol II in response to DNA damage. Assembly of the Elongin A ubiquitin ligase and its recruitment to sites of DNA damage is a tightly regulated process induced by DNA-damaging agents and α-amanitin, a drug that induces Pol II stalling. In this study, we demonstrate (i) that Elongin A and the ubiquitin ligase subunit CUL5 associate in cells with the Cockayne syndrome B (CSB) protein and (ii) that this interaction is also induced by DNA-damaging agents and α-amanitin. In addition, we present evidence that the CSB protein promotes stable recruitment of the Elongin A ubiquitin ligase to sites of DNA damage. Our findings are consistent with the model that the Elongin A ubiquitin ligase and the CSB protein function together in a common pathway in response to Pol II stalling and DNA damage.
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Juston C
authors.initials: JC
authors.lastname: Weems
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Slaughter
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Jay R
authors.initials: JR
authors.lastname: Unruh
authors.affiliation: the Mechanisms of Transcription Laboratory, The Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, United Kingdom.
authors.forename: Stefan
authors.initials: S
authors.lastname: Boeing
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Hall
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Merry B
authors.initials: MB
authors.lastname: McLaird
authors.affiliation: the Department of Functional Genomics, Kochi Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yasukawa
authors.affiliation: the Department of Functional Genomics, Kochi Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
authors.forename: Teijiro
authors.initials: T
authors.lastname: Aso
authors.affiliation: the Mechanisms of Transcription Laboratory, The Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, United Kingdom.
authors.forename: Jesper Q
authors.initials: JQ
authors.lastname: Svejstrup
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110, jlc@stowers.org.|the Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas 66160, and.
authors.forename: Joan W
authors.identifier: 0000-0002-2786-0663
authors.initials: JW
authors.lastname: Conaway
authors.affiliation: From the Stowers Institute for Medical Research, Kansas City, Missouri 64110, rcc@stowers.org.|the Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas 66160, and.
authors.forename: Ronald C
authors.initials: RC
authors.lastname: Conaway
mesh_terms: D053959:Alpha-Amanitin / Q000378:metabolism; D002460:Cell Line; D044842:Cullin Proteins / Q000378:metabolism; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism*; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000378:metabolism*; D000075065:Elongin; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D012319:RNA Polymerase II / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: D053959:Alpha-Amanitin; C496871:CUL5 protein, human; D044842:Cullin Proteins; C000616397:ELOA protein, human; D000075065:Elongin; D000075223:Poly-ADP-Ribose Binding Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D044767:Ubiquitin-Protein Ligases; D012319:RNA Polymerase II; D004265:DNA Helicases; C078673:ERCC6 protein, human; D045643:DNA Repair Enzymes
keywords: DNA repair; E3 ubiquitin ligase; RNA polymerase II; nucleotide excision repair; transcription