title: A specific GFP expression assay, penetrance estimate, and histological assessment for a putative splice site mutation in BRCA1.
abstract: Genetic testing for cancer predisposing mutations in BRCA1 and BRCA2 has been of benefit to many individuals from breast and ovarian cancer-prone kindreds. However, a function has not been assigned to many of the domains that make up these complex proteins and hence, the significance of many sequence variants, including missense mutations, splice-site mutations, and in-frame deletions/insertions, remains unclear. We identified a putative splice site mutation (IVS6-2delA) in BRCA1 in a family attending a Familial Cancer Centre that had a significant history of both breast and ovarian cancer. This sequence variant was not novel but the exact effect on mRNA splicing and hence the biological impact of this sequence variation was unclear and therefore the finding was unable to be used in genetic counseling of the family. Via the construction of novel GFP-based expression fusion constructs, we demonstrated that this sequence variation prevented normal splicing of the BRCA1 transcript. By combining these data with an assessment of the histopathological features of the breast carcinomas in this family and mutation penetrance estimate we were able to conclude that this BRCA1 variant conveyed an increased risk of breast cancer.
authors.affiliation: Department of Pathology, Peter MacCallum Cancer Institute, Melbourne, Australia. msouthey@unimelb.edu.au
authors.forename: M C
authors.initials: MC
authors.lastname: Southey
authors.forename: A
authors.initials: A
authors.lastname: Tesoriero
authors.forename: M A
authors.initials: MA
authors.lastname: Young
authors.forename: A J
authors.initials: AJ
authors.lastname: Holloway
authors.forename: M A
authors.initials: MA
authors.lastname: Jenkins
authors.forename: J
authors.initials: J
authors.lastname: Whitty
authors.forename: S
authors.initials: S
authors.lastname: Misfud
authors.lastname: kConFab
authors.forename: S A
authors.initials: SA
authors.lastname: McLachlan
authors.forename: D J
authors.initials: DJ
authors.lastname: Venter
authors.forename: J E
authors.initials: JE
authors.lastname: Armes
mesh_terms: D000328:Adult; D000368:Aged; D017398:Alternative Splicing / Q000235:genetics; D019313:BRCA1 Protein / Q000235:genetics*; D024682:BRCA2 Protein / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D019398:Genes, BRCA1; D006580:Genetic Carrier Screening / Q000379:methods; D020022:Genetic Predisposition to Disease / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010051:Ovarian Neoplasms / Q000235:genetics; D010375:Pedigree; D019683:Penetrance; D022821:RNA Splice Sites / Q000235:genetics*; D012334:RNA, Neoplasm / Q000032:analysis
chemical_list: D019313:BRCA1 Protein; D024682:BRCA2 Protein; D018076:DNA, Complementary; D008164:Luminescent Proteins; D022821:RNA Splice Sites; D012334:RNA, Neoplasm; D049452:Green Fluorescent Proteins