Chromosome loss with concomitant duplication and recombination both contribute most to loss of heterozygosity in vitro.
Loss of heterozygosity (LOH) plays an important role in the expression of recessive mutations in mammalian cells. To gain insight into the rate and mechanisms of LOH the autosomal HLA-A gene was used as a model system. Spontaneous HLA-A2 mutants originated with a rate of respectively 4.1 x 10(-6) and 6.9 x 10(-6) per cell per generation in TK6 and WI-L2-NS, two isogenic lymphoblastoid cell lines which differ in TP53 status. The rate of loss of HLA-A2 is 10-50 times higher compared to the mutation rate of the X-linked HPRT gene. The homozygous TP53 mutation in WI-L2-NS had no effect on the rate of HLA-A2 loss or the spectrum of these mutations. Microsatellite analysis of most of the HLA-A2 mutants (84%) showed LOH for multiple markers on chromosome arm 6p telomeric of a recombination breakpoint, LOH for all 6p markers, or LOH for markers on both the 6p- and 6q-arms. Cytogenetic analysis showed that these mechanisms gave mutant cells which harbored two intact chromosomes 6 and which were indistinguishable from non-mutant cells. Therefore, loss of HLA-A2 is mainly caused by somatic recombination (33-50%) or chromosome loss with duplication of the remaining chromosome (34-40%). These findings correspond to the mechanisms behind loss of the wild-type RBI allele in retinoblastoma and suggest that both somatic recombination and chromosome loss followed by duplication contribute to tumorigenesis.
de Nooij-van Dalen
A G
AG
MGC-Department of Radiation Genetics and Chemical Mutagenesis, Sylvius Laboratories, Leiden University, The Netherlands.
van Buuren-van Seggelen
V H
VH
Lohman
P H
PH
Giphart-Gassler
M
M
D002461:Cell Line, Transformed; D002872:Chromosome Deletion; D002896:Chromosomes, Human, Pair 6 / Q000235:genetics; D015234:HLA-A Antigens / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D019656:Loss of Heterozygosity; D011995:Recombination, Genetic
D015234:HLA-A Antigens