University of Leicester

was unearthed beneath a parking lot in Leicester, England. Based on the evidence at hand, the archeologists behind the discovery claimed that they had found the bones of Richard III—the 15th century monarch with the cartoonishly villainous reputation. (Richard, immortalized in one Shakespeare's greatest plays, reportedly stole the throne, killed his nephews, and ruled for two years before dying in battle.)

The evidence was compelling: The skeleton was male, the right age, from the right time period, had numerous battle injuries, and even showed Richard III's characteristic uneven shoulders and scoliosis. And good evidence had led the archaeologists to the parking lot, which is the site of a ruined monastery said to be the monarch's final resting place. Even so, the scientists couldn't be entirely sure it was really him—at least not without DNA evidence.

Today, a team of biologists, historians, and archeologists announced that they have completed a DNA analysis of the skeleton. As they report in the journal Nature Communications, these remains truly are the former king beyond any reasonable doubt. What's even more impressive: By tracing the genetic evidence through more than 20 generations of the dead king's family tree to his living descendants, the scientists have completed the oldest DNA identification case of a known individual.

"The evidence is overwhelming that these are indeed the remains of Richard III," says Turi King, the University of Leicester geneticist who led the DNA analysis, "Even with our most conservative statistical analysis, we're 99.999 percent sure."

Images: University of Leicester.

Genes Through Generations

King's team first had to scrape together a DNA profile from the withered skeleton—no easy task. Because DNA starts to fragment immediately after death, the researchers had to painstakingly sift through millions of ribbons of splintered DNA strands. To make sure the analysis wasn't contaminated (an ever-present risk), scientists did the sequencing in several clean rooms in universities across the U.K.

DNA profile in hand, the researchers compared it against living relatives from Richard III's family tree. Luckily, Richard's descendants are proud of their heritage—though the scientists had plenty of work to do finding the kind's real descendants and insulating the research from false claims, says Kevin Schürer, the chief University of Leicester historian with the research team.

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Also, King says, "not just any relative would do. We needed to find people who were related in a particular way. The vast majority of your DNA is a very complex mixture of your ancestors," she says, and an ancestor from 20 generations back would contribute just 1 millionth (on average) of your total DNA. That doesn't tell you a lot. "But there are two pieces of DNA that are passed down virtually unchanged through generations. That's your mitochondrial DNA," she says, "and the Y chromosome."

The mitochondria—the tiny powerhouse structure in all of your cells—have their own DNA. Mitochondrial DNA is passed on only by the mother. As a result, the scientists could compare their DNA analysis of the skeletal remains against two descendants by following an all-female line through their vetted family tree (from Richard III's mother all the way down to two living descendants.)

The female-line descendants shared the exact same mitochondrial DNA (except for a single mutation acquired through the generations) as Richard III. And what's more, it was also a rare variant of the DNA, one commonly found around the Mediterranean but rare in England.

Close-up of skeleton in the burial site.

Adultery Muddles Science Yet Again

For the Y chromosome—found only in men and passed down by the father—the scientists had a bit more trouble. Because Richard III has no pure male-line descendants in his family tree (his sons were violently murdered) the scientists and historians had to follow Richard III's Y chromosome back up his family tree to the 14th century king Edward III, before tracing an all-male line down. The scientists then identified five descendants—all of whom distantly shared their closest heritage through the former 5th Duke of Beaufort, Henry Somerset, who died in 1803.

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Here's where the story gets juicy. Only four of the five living descendants shared their Y chromosome. The 5th descendant either was either not a genetic descendant of the duke because of hidden adultery, or somewhere along the line someone had fabricated a fake claim of royal heritage. And while all four of Somerset's descendants shared a Y chromosome with each other, they did not share one with the remains of Richard the III.

After all this genetic sleuthing, this picture became clear: somewhere in the line of the family tree that extends up from Duke Henry Somerset to Edward III and down to Richard III, a queen or heiress became pregnant through adultery and then lied about it without anybody ever knowing.

It seems scientists have accidentally stumbled upon century-old royal infidelity—and crushed the royal claims of a few descendants. Hey, you never know what's going to happen when you go digging around in a parking lot.

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