Authors: Jo Marchant
Eleven mummies were included in the project. The first nine are familiar: Tutankhamun, the mummy from KV55, Amenhotep III, Yuya, Tjuiu, the two fetuses from Tutankhamun’s tomb, and the Elder and Younger Ladies from KV35. Added into the mix were two female mummies from a small nonroyal tomb called KV21. Pottery found in this tomb suggested that it dated from earlier in the Eighteenth Dynasty, and most Egyptologists assumed that the two unidentified women inside were noblewomen or courtiers. But they had bent left arms, so Hawass felt they could be relatives of Tutankhamun.
The plan was to compare the DNA of all these mummies to come up with a family tree. The researchers hoped that this would finally solve mysteries such as the identity of the KV55 mummy, Tutankhamun’s parents, and the fetuses from his tomb. The team would also test the mummies for DNA from the microbes that cause plague, tuberculosis, malaria, leishmaniasis, and leprosy, to see if they suffered from any of these diseases.
Meanwhile, Selim and his team continued CT scanning all of the mummies, and reanalyzed the scans of Tutankhamun. They were looking for any inherited characteristics or disorders that might run in the family, and of course, clues to how the various individuals died.
Gad and his team were testing for several different types of DNA. They were trying to amplify fragments of mitochondrial DNA (which is passed down the maternal line), as well as DNA from the male-determining Y chromosome, which is inherited from father to son.
But the main plan was to do genetic fingerprinting on the DNA that is passed down from both parents. This is a good way to look at family relationships without having to sequence the whole genome. We share almost all of our DNA with every other person on the planet; it’s what makes us human as opposed to chimps or grapes. But there are some small variations, which can be used to identify us.
DNA fingerprinting homes in on one specific type of variable region called microsatellites. If you look at enough different microsatellites, you end up with a “fingerprint” that’s unique for that particular person. In the royal mummy study, Zink and the team looked at eight microsatellites across the genome, with unwelcoming names like D13S317 and CSF1PO (which always makes me think of C3PO, the camp robot in Star Wars). You can use the details of a fingerprint to draw conclusions about family relationships, because we inherit our microsatellites (as the rest of our DNA) from our parents—half from one and half from the other.
Working conditions in the ancient DNA lab were now quite different from the Hatshepsut project in several respects. This time, the cameras had to stay outside. Hawass banned them after complaints from the team that the film crew was contaminating the lab—much to the annoyance of the Discovery producers, who after all had paid for the entire setup so that they could film the work done inside. Zink and Pusch also insisted that the researchers were given enough time to do their work properly.
And they took months. One problem, particularly for Tutankhamun, was that black resins and other materials used in the embalming process had crept right into the mummy’s bones. Normally, when you purify DNA from a sample, you end up with a clear solution. But to Zink and Pusch’s dismay, in Tutankhamun’s case, the samples turned out inky black. They couldn’t get rid of the impurities, which clung tight to the king’s DNA, blocking any subsequent reactions. According to Zink, it took six months of hard work to figure out how to remove the contaminants, but finally the precious DNA was ready for amplifying and sequencing.*
In Discovery’s film about the project,1 there’s a scene in which Zink and Pusch scrutinize a row of colored peaks on a computer screen. There is a dramatic pause. “My God!” whispers Pusch, the muffled words just audible from behind his surgical mask. Then the two hug and shake hands, accompanied by the laughter and applause of Gad and his Egyptian colleagues. The geneticists have just achieved their first successful result.
Except that they haven’t. As cameras weren’t allowed in the lab, the real moment of discovery wasn’t captured on film. The researchers were asked to stage a reenactment, weeks later, for the cameras. But they insist that their excitement in the dummy scene is real. The first DNA to show up from the mummies was from the Y chromosome. It showed that Amenhotep III, KV55, and Tutankhamun all belonged to the same male line, whereas an unrelated male mummy,* included as a control, did not. It was an important sign that the analysis was working well.
After that, the team moved on to the more challenging task of genetic fingerprinting. According to Zink, they painstakingly repeated the test twenty to thirty times on each mummy. Then they used a computer program to compare all of the fingerprints and come up with the all-important family tree.
Although popular with TV viewers, Hawass’s previous mummy studies had met with some skepticism among academics, partly because they weren’t first published in academic journals, meaning that other scientists had no chance to scrutinize and check the results. He didn’t want any such doubts with this study: Discovery would have to wait until the results were properly published. On February 17, 2010, the team published a ten-page paper in the prestigious Journal of the American Medical Association (JAMA).2 The same day, Hawass held a triumphant press conference at the Cairo museum. Gad, Selim, Hawass, and Pusch sat in a row to announce the results, their heads just visible over a forest of microphones carrying logos from TV companies all over the world.
What they had to say was dramatic, and appeared to change our historical understanding of the period. After decades of inconclusive results, the team at last offered some certainty. The study answered questions about Tutankhamun and his ancestry that had foxed archaeologists for decades, identified several anonymous mummies as being famous figures from the Amarna period, and completely changed our view of the boy king. The journalists loved it. But scientists watching from around the world weren’t so sure.
Impressively, the researchers said they had isolated DNA from every single mummy they tested. Seven mummies yielded a complete fingerprint across all eight microsatellites, while the two anonymous mummies from tomb KV21, and the two fetuses, produced partial data sets. Zink and Pusch used these results to construct a five-generation family tree, with the elderly couple Yuya and Tjuiu at the top. The DNA appeared to confirm that the Elder Lady was indeed Yuya and Tjuiu’s daughter Queen Tiye, as originally suggested by James Harris, and supported by Selim’s CT scans. (I can’t help wondering if perhaps Pusch was just a tiny bit influenced by the mummy’s striking looks, when he told journalists that her genetic material was “the most beautiful DNA that I’ve ever seen from an ancient specimen.”3)
The team concluded that the troublesome KV55 mummy was the son of Amenhotep III and Tiye—most probably Akhenaten—and the father of Tutankhamun. The computer analysis seemed unarguable. The calculated probability that the KV55 mummy was Tutankhamun’s father, for example, was a slam-dunk 99.99999981 percent.
The DNA analysis seemed to confirm Selim’s controversial suggestion that the Younger Lady from tomb KV35 was Tutankhamun’s mother. And to the great surprise of pretty much every Egyptologist with an interest in the period, she appeared to be another—previously unknown—child of Amenhotep III and Tiye. In other words, Tutankhamun’s parents were brother and sister.
The incomplete data on the fetuses weren’t enough for a definitive identification but supported the idea that they were indeed Tutankhamun’s stillborn daughters. And there was another surprise: the partial DNA profile of one of the mummies found in tomb KV21 suggested that she could be their mother—presumably Tutankhamun’s wife Ankhesenamun. If Egyptologists were surprised to find Tiye and Tutankhamun’s mother in a side room of Amenhotep II’s tomb, the idea that Ankhesenamun had been lying unnoticed in KV21 seemed even weirder, because there was nothing else in the tomb to hint that it had anything to do with Tutankhamun’s reign. It seemed an incredible stroke of luck that Hawass had included these unpromising mummies in the study.
Most of the tests for disease DNA came out negative. But the researchers did detect DNA from Plasmodium falciparum, the parasite that causes the most virulent form of malaria, in Yuya, Tjuiu, and Tutankhamun.
Selim presented the latest results of the CT scans. Surprisingly, there was plenty new to say about Tutankhamun, even though this study had already been reported once. In fact, much of the updated analysis contradicted conclusions that the team had reached the first time around.
Rather than the well-fed, healthy youth suggested by the initial study, Selim now argued that the young king was riddled with minor ailments and weaknesses. For example, Selim diagnosed scoliosis of the spine in Tutankhamun—as well as in his mother, grandmother, and two stillborn children. What previous investigators had agreed was simply a result of how Tutankhamun’s body was laid out by his embalmers was now an inherited disorder affecting almost every member of the family.
Selim also concluded that Tutankhamun had a badly deformed left foot. As well as a clubfoot, the accompanying JAMA paper reported, the king had been born with a missing toe bone, and one of his metatarsals was being eaten away by necrosis, a disease in which a lack of blood supply causes the bone tissue to die. The team diagnosed clubfoot in KV55 and the two KV21 mummies too, and reported (among other things) that Yuya, Tjuiu, and Tiye were affected by festering surgical wounds.
In summary, the royal family was a sickly bunch, with Tutankhamun in the worst shape of all—a frail, limping cripple who needed a stick to walk. He was hardly the type you might expect to have met his end falling from a speeding chariot, as had been suggested in 2005.
The one thing Tutankhamun didn’t suffer from, apparently, was a gender disorder. This might sound a bit random, but Egyptologists and medics alike had been speculating about this for decades. The debate was triggered mostly by those bizarrely curvaceous statues of Akhenaten, but there are some rather feminine figures of Tutankhamun, too.
Some experts think this was just part of the Amarna art revolution, with Akhenaten determined to show himself (and perhaps his family members) as strikingly different from their predecessors, with attributes of both the father and mother of all creation. But many scholars have seen evidence of an inherited disorder, passed from father down to son.
Speculation about Akhenaten’s medical condition started early, soon after his strange statues were discovered. In 1855, Auguste Mariette, founder of the antiquities service, suggested that the king had been taken prisoner in Sudan and castrated. Another French Egyptologist, Gustave Lefebvre, wondered in 1890 if he was a woman masquerading as a man.
In 1907, the anatomist Grafton Elliot Smith accredited the pharaoh’s peculiar physique to Fröhlich syndrome: a rare childhood metabolic disorder characterized by obesity, retarded growth, and delayed development of genital organs. But the syndrome also causes infertility, and it was hard to reconcile this with the fact that Akhenaten had apparently fathered six daughters (and perhaps some sons too). The Egyptologist Cyril Aldred subsequently suggested in the 1960s that perhaps Akhenaten was born normal, allowing him to father children, but later suffered from a tumor affecting his pituitary gland, which caused his symptoms later in life.4
In the 1970s, British medics focused on Tutankhamun himself, after seeing gilded figures of the king in the British Museum’s touring exhibition that showed him with a sagging tummy and pert breasts.* In a series of letters to top medical journal The Lancet, various experts diagnosed poor Tutankhamun with a dizzying range of conditions, including an adrenal tumor; Klinefelter’s syndrome (in which men carry an extra X chromosome in their cells); Wilson’s disease (in which there’s too much copper in the body’s tissues); or even a “Tutankhamun syndrome” new to medical science.6 Eventually Ronald Harrison felt obliged to write in, pointing out that as the only expert who had actually studied Tutankhamun’s mummy, as opposed to simply admiring statues in an exhibition, he could confirm that there was no evidence the king had any of the suggested disorders.7
That didn’t stop the speculation though. In 1980, a doctor from Philadelphia named Bernadine Paulshock suggested that because certain statues of Amenhotep III, Akhenaten, Smenkhkare, and Tutankhamun all show signs of breasts, this could represent a genetic disorder running in the family.8 She suggested pseudohermaphroditism, where males also exhibit female characteristics, a condition that she argued was common in families with a long history of inbreeding.
In the 1990s, Bob Brier suggested that Akhenaten, with his elongated head and spindly limbs, suffered from Marfan syndrome, a disorder of the connective tissue that holds muscles and bones together.9 And in 2009, scientists from Yale University came up with yet another suspect: a hormonal disorder called Antley-Bixler syndrome, in which a single mutated gene causes an elongated skull as well as overproduction of the sex hormone estrogen.10
Selim and his colleagues looked for signs of some of these disorders, but like Harrison found no evidence that either Tutankhamun or the KV55 mummy suffered any such problems. Their skulls, although very wide, weren’t pathological. Tutankhamun’s penis, albeit detached from his body, looked normal too. They concluded that the womanly appearance of the statues was probably “a royally decreed style,” and that neither Tutankhamun nor Akhenaten actually had a significantly bizarre or feminine physique.
To summarize, then, Tutankhamun was a full-blown male, at least. But he was no longer an active youth who perhaps died in a hunting accident. Instead he was a weak, inbred cripple, afflicted by a series of congenital disorders. Oh, and he was infected with malaria.
As supporting evidence for the king’s diseased state, the team pointed to various foodstuffs found in Tutankhamun’s tomb, including fruits, seeds, and oils that had possible medicinal uses. For example, the tomb contained thirty-six baskets of date-like fruit from the Zizyphus or Christ’s-thorn tree, which was primarily eaten as food, but also prescribed to treat stomach complaints, fever, and pain. Coriander seeds, found in eight baskets, could be used to treat gastrointestinal complaints, infected wounds, or “demonic” diseases of the head.