Processes of de novo duplication of human α-globin genes

Ectopic recombination between repeated but nonallelic DNA sequences plays a major role in genome evolution, creating gene families and generating copy number variation and pathological rearrangements in human chromosomes. Previous studies on the alpha 2- and alpha 1-globin genes have shown that de novo deletions common in alpha(+)-thalassemics can be directly accessed in human DNA and provide an informative system for studying deletion dynamics and processes. However, nothing is known about the reciprocal products of ectopic recombination, namely gene duplications. We now show that molecules carrying three alpha-globin genes can be detected in human DNA by using physical enrichment plus an inverse PCR strategy. These de novo duplications are common in blood and sperm and appear to arise by two distinct mechanisms: meiotic exchanges between homologous chromosomes that generate a minority of sperm duplications, plus mitotic ectopic exchanges that occur in the soma and germ line and can show erratic fluctuations in frequency most likely caused by mutational mosaicism. The dynamics and processes of duplication are very similar to those of deletion, particularly for meiotic exchanges. This result suggests rearrangement pathways dominated by fully reciprocal ectopic exchange, with nonreciprocal pathways such as intramolecular recombination and single-strand annealing playing at best only a minor role in the generation of deletions. Finally, the high level of instability at the a-globin locus contrasts with the rarity in most populations of chromosomes carrying duplications or deletions, pointing to strong selective constraints that maintain alpha-globin gene copy number in human populations.