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Despite his brief nine-year reign, Tutankhamen is probably the most famous pharaoh of ancient Egypt. Because his tomb had not been robbed at the time of its discovery in 1922, historians have been able to piece together aspects of the boy king's 19-year life. More than 100 walking sticks and "pharmacies" (medicinal seeds, fruits and leaves) found mingled among funeral offerings and other treasures within the tomb suggested that the pharaoh was frail, and two mummified fetuses implied that his offspring might have suffered from lethal genetic defects. But a new study on the Tutankhamen family mummies themselves, published February 16 in JAMA The Journal of the American Medical Association, has provided biological insight into the king's incestuous royal lineage and his early death.
Secretary General Zahi Hawass of Egypt's Supreme Council of Antiquities teamed up with paleogeneticist Carsten Pusch from the University of Tübingen in Germany, to examine Tutankhamen and 10 royal mummies, including the two fetuses, presumed to be related to him for kinship, inherited disorders and infectious diseases. Five mummies that were thought to be unrelated served as morphological and genetic controls. Hawass, Pusch and 15 other scientists continue to perform detailed anthropological, radiological and genetic studies on the precious mummies in a lab built into the basement of the Museum of Egyptian Antiquities in Cairo—two floors below the famous golden mask.
After extracting tiny amounts of ancient DNA from the mummies' bones, the researchers amplified 16 short tandem repeats (short sequences in the DNA that create a genetic fingerprint) and eight polymorphic microsatellites (hereditary molecular markers) to testable quantities using techniques commonly employed in criminal or paternity investigations. They also looked for DNA sequences from the malaria pathogen.
Based on their results so far, the researchers were able to name several mummies who were previously anonymous (referred to only by tomb number), including Tut's grandmother "Tiye" and Tut's father, the KV55 mummy probably named "Akhenaten". "This is the most important discovery since the finding of the tomb of Tutankhamen in 1922," Hawass says. The team also identified the mummy likely to be Tut's mother as KV35YL, not Queen Nefertiti as was once thought. "Now I'm sure that it cannot be Nefertiti, and therefore the mother of King Tut is one of the daughters of Amenhotep III and Tiye—and there are five," Hawass says, adding that he plans to investigate this further.
"The more data we collected, the more the museum specimens came back to life," Pusch says, who admits he was worried about working with such ancient "pharaohic" DNA. "We had 16 mummies," Pusch explained. "You have a lot to do in the lab when you have a single mummy!" But the embalming process used to preserve these royal remains worked in Pusch's favor. "Some embalming really enhances the preservation of genetic material," he says.
The study revealed that King Tut's parents were siblings, a trend which might have continued in Tut's marriage. "There are rumors that Tut's wife was his sister or half sister. If this is true we have at least two successive generations that had interfamilial marriages, and this is not a good thing," Pusch says. "We see it quite often in royal families that they marry each other. They thought: 'Better to stay close.' I think we cannot judge from the insight we have nowadays." Because only partial DNA was retrieved from the mummified fetuses, it is still unclear whether they were Tut's offspring or just ceremonial offerings.
In 2005 Hawass performed a computed tomography (CT) scan to determine the cause of the boy pharaoh's death. The scan revealed a fractured femur, which could have caused death from infection or from a blood clot. The present study revealed that juvenile aseptic bone necrosis—a disorder in which poor blood supply leads to bone damage—might have rendered Tut particularly vulnerable to physical injuries. "We know that this man had 130 walking sticks and that he used to shoot arrows while he was sitting," Hawass says.
But the genetic analysis identified DNA from the malaria tropica (Plasmodium falciparum) pathogen, suggesting that Tut was also hampered by infection. "Unfortunately this was the worst form of malaria. Even today we don't have very good medications to deal with it," Pusch explains. The team concluded that a sudden leg fracture might have led to a life-threatening condition when a malaria infection occurred. "He was not a proud pharaoh or a strong leader, he was a young boy—frail and weak. He couldn't walk by himself and needed other people or walking sticks because of this bone necrosis," Pusch says, explaining that Tutankhamen's family was plagued by malformations and infections. "When I was a boy, I thought, 'Wow, these are royals and queens,' but they were suffering. They had pain, chills and fevers."
The study is the first to examine DNA from King Tut and his royal family, according to the researchers. "It's really a new kind of scientific discipline," Pusch says, calling it molecular anthropology. (The paper refers to it as molecular Egyptology.) "You have to be careful with these things. It's about evolution and history. The more you examine the mummy, the smaller it will become."
Pusch says he is grateful to have been a part of the project, adding that he couldn't sleep for three days after first arriving in the lab below the museum. "I remember going there with my parents when I was 12. To go back 30 years later to work with the original one and only, sometimes I wake up and think it's a dream!" Pusch says that the project has answered many questions but raised many more. "We have so many exciting questions for the future. In the meantime, we’ll gather more and more questions—it will never end!"