The discovery of introns and the split gene architecture of the eukaryotic genes started a new era of eukaryotic biology. The question of why eukaryotic genes had fragmented genes prompted speculation and discussion almost immediately.
Ford Doolittle published a paper in 1978 in which he stated that most molecular biologists assumed that the eukaryotic genome arose from a ‘simpler’ and more ‘primitive’ prokaryotic genome rather like that of Escherichia coli. However, this type of evolution would require that introns be introduced into the coding sequences of bacterial genes. Regarding this requirement, Doolittle said, “It is extraordinarily difficult to imagine how informationally irrelevant sequences could be introduced into pre-existing structural genes without deleterious effects.” He stated “I would like to argue that the eukaryotic genome, at least in that aspect of its structure manifested as ‘genes in pieces’ is in fact the primitive original form.”
James Darnell expressed similar views in 1978. He stated, “The differences in the biochemistry of messenger RNA formation in eukaryotes compared to prokaryotes are so profound as to suggest that sequential prokaryotic to eukaryotic cell evolution seems unlikely. The recently discovered non-contiguous sequences in eukaryotic DNA that encode messenger RNA may reflect an ancient, rather than a new, distribution of information in DNA and that eukaryotes evolved independently of prokaryotes.”
However, in an apparent attempt to reconcile with the idea that RNA preceded DNA in evolution, and with the concept of the three evolutionary lineages of archea, bacteria and eukarya, both Doolittle and Darnell deviated from their original speculation in a joint paper in 1985. They suggested that the ancestor of all three groups of organisms, the ‘progenote,’ had a genes-in-pieces structure, from which all three lineages evolved. They speculated that the precellular stage had primitive RNA genes which had introns, which were reverse transcribed into DNA and formed the progenote. Bacteria and archea evolved from the progenote by losing introns, and ‘urkaryote’ evolved from it by retaining introns. Later, the eukaryote evolved from the urkaryote by evolving a nucleus and absorbing mitochondria from bacteria. Multicellular organisms then evolved from the eukaryote.
These authors predicted that the distinctions between the prokaryote and the eukaryote were so profound that the prokaryote to eukaryote evolution was not tenable, and had different origins. However, other than the speculations that the precellular RNA genes must have had introns, they did not address the key questions of intron origin. No explanations described why exons were short and introns were long, how the splice junctions originated, what the structure and sequence of the splice junctions meant, and why eukaryote genomes were large.
Around the same time that Doolittle and Darnell suggested that introns in eukaryotic genes could be ancient, Colin Blake and Walter Gilbert published their views on intron origins independently. In their view, introns originated as spacer sequences that enabled convenient recombination and shuffling of exons that encoded distinct functional domains in order to evolve new genes. Thus, new genes were assembled from exon modules that coded for functional domains, folding regions, or structural elements from preexisting genes in the genome of an ancestral organism, thereby evolving genes with new functions. They did not specify how exons or introns originated. In addition, even after many years, extensive analysis of thousands of proteins and genes showed that only extremely rarely do genes exhibit the supposed exon shuffling phenomenon. Furthermore, molecular biologists questioned the exon shuffling proposal, from a purely evolutionary view for both methodological and conceptual reasons, and, in the long run, this theory did not survive.