needed to activate the receptor goes up with each step in the left-hand box, and is even higher ineach animal in the right-hand box. It takes more hormone to activate the receptor – it became lesssensitive, and seems to have acquired this characteristic a long time ago.How long ago, and how would did the authors infer that? Well, they created a postulatedreconstruction of the ancestral receptor – the receptor in the common ancestor – by combiningknowledge of rates of change in proteins with the new knowledge gained by looking at four newanimals (the sharks). The graph in Figure 2 shows the hormone sensitivity of this new reconstruction(it's version 1.1) compared to an older reconstruction based on just one cartilaginous sh (that wasversion 1.0). The new "resurrected" ancestral receptor (AncGR1.1) has much lower sensitivity thanits much more ancient ancestor (AncCR), the one with high sensitivity but no selectivity. So it seemsthat the change in sensitivity happened after the duplication event but before any of the various kindsof vertebrates diverged, something less than 450 million years ago.But
did the change come about? Let's look back at the family tree in Figure 1 to get oriented.The grandparent of all the receptors is called AncCR and it was sensitive but not selective. After theduplication, there were two parents, if you will: the MR parent which we're not discussing, and theGR parent, called the AncGR. AncGR, the parent of
the GRs, had reduced sensitivity. Carroll andcolleagues addressed the
question by rst looking at the types of changes that occurred betweenthe grandparent and the parent.There were 36 changes that accrued during that time. The authors used some straightforwardreasoning to narrow the list of suspects down to six. In other words, six different changes in theprotein, together or separately, were likely to account for the change in sensitivity. They went intothe lab and resurrected each of those mutant proteins, and measured their sensitivity. And their datatell a very interesting story, presented as a graph in
.The gray bar is the grandparent. The yellow bar is the parent. (The scale is logarithmic, so thechange in sensitivity from grandparent to parent is at least 100-fold.) The other bars represent thesensitivity of some of the mutations that must have generated the low-sensitivity parent. So, the rstwhite bar is mutant 43. (The number represents a particular location of the mutation, but we're notinterested in that here.) That mutation drops sensitivity to near-parental levels. The next bar ismutant 116. It also reduces sensitivity to the parental level. Sounds good. But wait:
of thosemutations are present in the parent. What happens when you put them together? Disaster. Look at thenext white bar: it's the combination of 43 and 116, and the receptor is effectively dead. As theauthors put it in the abstract, "the degenerative effect of these two mutations is extremely strong."