title: XPC branch-point sequence mutations disrupt U2 snRNP binding, resulting in abnormal pre-mRNA splicing in xeroderma pigmentosum patients.
abstract: Mutations in two branch-point sequences (BPS) in intron 3 of the XPC DNA repair gene affect pre-mRNA splicing in association with xeroderma pigmentosum (XP) with many skin cancers (XP101TMA) or no skin cancer (XP72TMA), respectively. To investigate the mechanism of these abnormalities we now report that transfection of minigenes with these mutations revealed abnormal XPC pre-mRNA splicing that mimicked pre-mRNA splicing in the patients' cells. DNA oligonucleotide-directed RNase H digestion demonstrated that mutations in these BPS disrupt U2 snRNP-BPS interaction. XP101TMA cells had no detectable XPC protein but XP72TMA had 29% of normal levels. A small amount of XPC protein was detected at sites of localized ultraviolet (UV)-damaged DNA in XP72TMA cells which then recruited other nucleotide excision repair (NER) proteins. In contrast, XP101TMA cells had no detectable recruitment of XPC or other NER proteins. Post-UV survival and photoproduct assays revealed greater reduction in DNA repair in XP101TMA cells than in XP72TMA. Thus mutations in XPC BPS resulted in disruption of U2 snRNP-BPS interaction leading to abnormal pre-mRNA splicing and reduced XPC protein. At the cellular level these changes were associated with features of reduced DNA repair including diminished NER protein recruitment, reduced post-UV survival and impaired photoproduct removal.
authors: Dermatology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland 20892, USA.
authors: Sikandar G
authors: SG
authors: Khan
authors: Koji
authors: K
authors: Yamanegi
authors: Zhi-Ming
authors: ZM
authors: Zheng
authors: Jennifer
authors: J
authors: Boyle
authors: Kyoko
authors: K
authors: Imoto
authors: Kyu-Seon
authors: KS
authors: Oh
authors: Carl C
authors: CC
authors: Baker
authors: Engin
authors: E
authors: Gozukara
authors: Ahmet
authors: A
authors: Metin
authors: Kenneth H
authors: KH
authors: Kraemer
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics*; D005091:Exons / Q000235:genetics; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D011485:Protein Binding / Q000528:radiation effects; D011740:Pyrimidine Dimers / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics* / Q000528:radiation effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000378:metabolism*; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics* / Q000473:pathology
chemical_list: D004268:DNA-Binding Proteins; D011740:Pyrimidine Dimers; D012322:RNA Precursors; D012333:RNA, Messenger; D017413:Ribonucleoprotein, U2 Small Nuclear; C086984:XPC protein, human; D004247:DNA; D016914:Ribonuclease H