Catalytically impaired hMYH and NEIL1 mutant proteins identified in patients with primary sclerosing cholangitis and cholangiocarcinoma.
The human hMYH and NEIL1 genes encode DNA glycosylases involved in repair of oxidative base damage and mutations in these genes are associated with certain cancers. Primary sclerosing cholangitis (PSC), a chronic cholestatic liver disease characterized by inflammatory destruction of the biliary tree, is often complicated by the development of cholangiocarcinoma (CCA). Here, we aimed to investigate the influence of genetic variations in the hMYH and NEIL1 genes on risk of CCA in PSC patients. The hMYH and NEIL1 gene loci in addition to the DNA repair genes hOGG1, NTHL1 and NUDT1 were analyzed in 66 PSC patients (37 with CCA and 29 without cancer) by complete genomic sequencing of exons and adjacent intronic regions. Several single-nucleotide polymorphisms and mutations were identified and severe impairment of protein function was observed for three non-synonymous variants. The NEIL1 G83D mutant was dysfunctional for the major oxidation products 7,8-dihydro-8-oxoguanine (8oxoG), thymine glycol and dihydrothymine in duplex DNA, and the ability to perform delta-elimination at abasic sites was significantly reduced. The hMYH R260Q mutant had severe defect in adenine DNA glycosylase activity, whereas hMYH H434D could excise adenines from A:8oxoG pairs but not from A:G mispairs. We found no overall associations between the 18 identified variants and susceptibility to CCA in PSC patients; however, the impaired variants may be of significance for carcinogenesis in general. Our findings demonstrate the importance of complete resequencing of selected candidate genes in order to identify rare genetic variants and their possible contribution to individual susceptibility to cancer development.
Forsbring
Monika
M
Institute of Medical Microbiology, Rikshospitalet University Hospital HF and Centre for Molecular Biology and Neuroscience, University of Oslo, Oslo, Norway
Vik
Erik S
ES
Dalhus
Bjørn
B
Karlsen
Tom H
TH
Bergquist
Annika
A
Schrumpf
Erik
E
Bjørås
Magnar
M
Boberg
Kirsten M
KM
Alseth
Ingrun
I
D001650:Bile Duct Neoplasms / Q000201:enzymology* / Q000235:genetics; D001653:Bile Ducts, Intrahepatic; D018281:Cholangiocarcinoma / Q000201:enzymology* / Q000235:genetics; D015209:Cholangitis, Sclerosing / Q000201:enzymology* / Q000235:genetics; D004249:DNA Damage; D045647:DNA Glycosylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005091:Exons; D005260:Female; D020022:Genetic Predisposition to Disease; D006801:Humans; D007438:Introns; D008297:Male; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D012306:Risk
D045647:DNA Glycosylases; C473634:NEIL1 protein, human; C061738:mutY adenine glycosylase