One basis for the evolution of organisms is the acquisition of new temporal and spatial domains of gene expression. Such novel expression domains could be generated either by cis sequence changes that alter the complement of trans-acting regulators binding to control elements or by changes in the expression patterns of one or more of the regulatory (trans) factors themselves. The ␥ globin gene is a prime example of a gene that has undergone a distinct change in temporal expression at a defined time in evolution. Approximately 35-55 million years ago, the previously embryonic ␥ gene acquired a fetal expression pattern. This change occurred in a simian primate ancestor after the separation of simian and prosimian primates but before the further separation of the major simian lineages; thus, the (prosimian) galago ␥ gene retains the ancestral embryonic expression pattern, whereas the (simian) human ␥ gene is fetal. This analysis of galago and human ␥ genes in transgenic mice demonstrates that cis changes in sequences within a 4.0-kilobase region surrounding the ␥ gene were responsible for the evolution of a novel fetal expression pattern in the ␥ globin genes of simian primates.

Stage-specific expression of the human β-like globin genes is controlled by interactions between regulatory elements near the individual genes and additional elements located upstream in the Locus Control Region (LCR). Elucidation of the mechanisms that govern these interactions could suggest strategies to reactivate fetal (γ) or embryonic (ɛ) genes in individuals with severe hemoglobinopathies. This study extends an earlier analysis of a transgenic construct, HS3ɛγ, testing: (A) the effect of substitution of HS2 for HS3 on stage-specific expression of the ɛ and γ genes and, (B) the role of an evolutionarily conserved YY1 binding site in transcriptional regulation of the γ gene. The data show that both HS3ɛγ and HS2ɛγ can individually support embryonic expression of ɛ and fetal expression of Aγ. Thus, the cis regulators of distinct expression patterns for ɛ and γ are likely to reside near the genes, rather than in specific hypersensitive sites of the LCR. Alterations in Aγ expression patterns observed in transgenic lines carrying a construct with a mutation in a conserved YY1 binding site at −1086 indicate that this site might function to facilitate active transcription of the γ gene in fetal life.