title: Assessment of human Nter and Cter BRCA1 mutations using growth and localization assays in yeast.
abstract: A large number of missense mutations have been identified within the tumor suppressor gene BRCA1. Most of them, called "variants of unknown significance" (VUS), cannot be classified as pathogenic or neutral by genetic methods, which complicates their cancer risk assessment. Functional assays have been developed to circumvent this uncertainty. They aim to determine how VUS impact the BRCA1 protein structure or function, thereby giving an indication of their potential to cause cancer. So far, three relevant assays have been designed in yeast and used on large sets of variants. However, they are limited to variants mapped in restricted domains of BRCA1. One of them, the small colony phenotype (SCP) assay, monitors the BRCA1-dependent growth of yeast colonies that increases with pathogenic but not neutral mutations positioned in the Cter region. Here, we extend this assay to the Nter part of BRCA1. We also designed a new assay, called the "yeast localization phenotype (YLP) assay," based on the accumulation of BRCA1 in a single inclusion body in the yeast nucleus. This phenotype is altered by variants positioned both in the Nter and Cter regions. Together, these assays provide new perspectives for the functional assessment of BRCA1 mutations in yeast.
authors.affiliation: Institut Curie, Centre de Recherche, 26 rue d'Ulm, Paris, France. gael.millot@curie.fr
authors.forename: Gaël A
authors.initials: GA
authors.lastname: Millot
authors.forename: Adeline
authors.initials: A
authors.lastname: Berger
authors.forename: Vincent
authors.initials: V
authors.lastname: Lejour
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Boulé
authors.forename: Claude
authors.initials: C
authors.lastname: Bobo
authors.forename: Christophe
authors.initials: C
authors.lastname: Cullin
authors.forename: Judith
authors.initials: J
authors.lastname: Lopes
authors.forename: Dominique
authors.initials: D
authors.lastname: Stoppa-Lyonnet
authors.forename: Alain
authors.initials: A
authors.lastname: Nicolas
mesh_terms: D019313:BRCA1 Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods*; D001943:Breast Neoplasms / Q000235:genetics / Q000503:physiopathology*; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure*; D005260:Female; D019398:Genes, BRCA1; D020022:Genetic Predisposition to Disease; D014644:Genetic Variation; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism*; D009154:Mutation; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
chemical_list: D019313:BRCA1 Protein