Gene amplification by unequal sister chromatid exchange: Probabilistic modeling and analysis of drug resistance data

Unequal sister chromatid exchange has been proposed as one of several possible mechanisms for gene amplification resulting in tandemly repeated sequences on chromosomes. Two requirements for testing this hypothesis are analytical observations and a mathematical model. Recently observations were reported for the number of tandemly repeated sequences on chromosomes of cells growing in the presence of a toxic drug and the mechanism was proposed to be unequal sister chromatid exchange. We now develop a mathematical model of this process based on the following hypotheses, (i) the extent of slippage between paired sister chromatids is a random variable with geometric distribution, (ii) the number of crossover sites is a random variable with a Poisson distribution, and (iii) cells with less than a threshold number of copies of an essential gene are eliminated when grown in selective conditions. Iterating the model at successive cell divisions results in a Markov chain with a denumerable infinity of states. The resulting distributions of gene copy number per cell at a particular population size are compared to published data on the CAD gene in BHK cells growing in the presence of the drug PALA (Smith et al., 1990, Cell, 63, 1219). The mathematical model can reproduce the observed means and standard deviations of gene copy number per cell and allows construction of confidence region estimates of parameters describing the extent of slippage, density of crossover sites, and strength of selection. An important prediction of the model is that in non-selective conditions the cells with amplified sequences gradually disappear from the population even if they are not at a growth disadvantage, though rare cells with a very large number of amplified sequences might continue to exist. The success of modeling suggests that the proposed mechanism of gene amplification by unequal sister chromatid exchange is consistent with the number of tandemly repeated sequences on chromosomes observed in some circumstances.