title: Trypanosoma brucei harbours a divergent XPB helicase paralogue that is specialized in nucleotide excision repair and conserved among kinetoplastid organisms.
abstract: Conserved from yeast to humans, TFIIH is essential for RNA polymerase II transcription and nucleotide excision repair (NER). TFIIH consists of a core that includes the DNA helicase Xeroderma pigmentosum B (XPB) and a kinase subcomplex. Trypanosoma brucei TFIIH harbours all core complex components and is indispensable for RNA polymerase II transcription of spliced leader RNA genes (SLRNAs). Kinetoplastid organisms, however, possess two highly divergent XPB paralogues with only the larger being identified as a TFIIH subunit in T. brucei. Here we show that a knockout of the gene for the smaller paralogue, termed XPB-R (R for repair) resulted in viable cultured trypanosomes that grew slower than normal. XPB-R depletion did not affect transcription in vivo or in vitro and XPB-R was not found to occupy the SLRNA promoter which assembles a RNA polymerase II transcription pre-initiation complex including TFIIH. However, XPB-R(-/-) cells were much less tolerant than wild-type cells to UV light- and cisplatin-induced DNA damage, which require NER. Since XPB-R(-/-) cells were not impaired in DNA base excision repair, XPB-R appears to function specifically in NER. Interestingly, several other protists possess highly divergent XPB paralogues suggesting that XPBs specialized in transcription or NER exist beyond the Kinetoplastida.
authors.affiliation: Department of Genetics and Developmental Biology, University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Nitika
authors.initials: N
authors.lastname: Badjatia
authors.forename: Tu N
authors.initials: TN
authors.lastname: Nguyen
authors.forename: Ju Huck
authors.initials: JH
authors.lastname: Lee
authors.forename: Arthur
authors.initials: A
authors.lastname: Günzl
mesh_terms: D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D019143:Evolution, Molecular; D055786:Gene Knockout Techniques; D017125:Genes, Protozoan; D006801:Humans; D016830:Kinetoplastida / Q000145:classification / Q000201:enzymology / Q000235:genetics; D010802:Phylogeny; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D051758:Transcription Factor TFIIH / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics*
chemical_list: D015800:Protozoan Proteins; D051758:Transcription Factor TFIIH; D004265:DNA Helicases