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Research in the News: DNA's Role - Tales From the Crypt (Grades 9-12)

Research in the News: Tales From the Crypt

Ruth Levy Guyer, Ph.D.

It was a gruesome end. On July 17, 1918, the Bolsheviks rounded up the members of Russia's Imperial Family, and 12 executioners shot the Czar, the Czarina, their children, the family doctor, and some servants. The bodies vanished and accounts of the events of the evening differed. The biggest puzzle was whether Anastasia, one of the four teen-aged daughters, had indeed died that night or had escaped. The uncertainties about the fate of the youngest girl in the family first surfaced in 1920, when a rescued, drowning woman in Berlin claimed that she was Anastasia.

The Russian Imperial Family. Taken from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson, Ltd., London.
The Russian Imperial Family. Taken from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson, Ltd., London

Surviving members of the Imperial family and their friends could not agree on whether the woman, Anna Anderson Manahan, was Anastasia (1). Some were sure she was; others said no. Manahan fought from 1938 to 1970 to reclaim what she insisted was her heritage. But, after 32 years of litigation, the verdict was still uncertain. Fourteen years later, in 1984, Manahan died (2).

The Four Grand Duchesses -- Marie, Tatiana, Anastasia, Olga.  Taken from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson Publishing, Ltd., London.
The Four Grand Duchesses -- Marie, Tatiana, Anastasia, Olga. Taken from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson Publishing, Ltd., London

That might have been the end of the story. But, in 1989, a document that had been written in 1920 was found. It described the night of the massacre and what happened to the bodies (2). When the shooting ended, some of the family and some of the servants were still alive. The executioners then stabbed those who were still breathing. They stripped the bodies and burned them with gasoline. Just to make future identification of the bones harder, they doused the bodies with sulfuric acid. The bones were thrown into a pit outside Ekaterinburg, and the pit was filled with dirt.

The report gave clues to the location of the grave, and, in 1991, two Russians -- an amateur historian and a former policeman who had turned to writing thrillers -- found what they figured might be the burial site. Government officials soon took over the investigation and pulled nine skeletons from the pit.

The Russians asked scientists in England to work with them to examine the bones. They speculated that newly available DNA technologies -- which involve extracting tiny amounts of DNA from bone fragments and then amplifying the DNA so that there is enough material to study -- might help in the identification of the bones and in determining their relations to each other. Were these the bones of the Imperial family? Could they shed light on Anastasia's fate? The findings were published in an article in Nature Genetics (3). The bones were evaluated with three different DNA tests.

The first involved identification of a gene that is found on both the X and the Y chromosomes. It is slightly different in the two and thus distinguishes male from female bones. This DNA test showed that there were five female and four male bodies in the grave.

Picture of an X and Y chromosome.
Picture of an X and Y chromosome

The next test assessed regions of DNA -- called short tandem repeats (STRs) -- in the samples. STR tests help in establishing whether individuals are related to one another: relatives have similar patterns and sets of STRs, just as they might share facial features.

The Czar and Alexei. Photograph from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson Publishing, Ltd., London.
The Czar and Alexei. Photograph from "The Last Days of Imperial Russia" by Miriam Kochan. Courtesy of Weidenfeld & Nicholson Publishing, Ltd., London

The STR tests showed that five of the skeletons in the grave were from people who were related -- they fit the profile of a family. Skeletons 4 and 7 were the parents and skeletons 3, 5, and 6 were their children. Sex testing had shown that 3, 5, and 6 were girls. Thus, the bones could be those of the Czar and Czarina and three of their four daughters. One daughter (Anastasia?) was missing and so was their son. What happened to Anastasia and Alexei? They might have been buried elsewhere; they might have survived.

A cell with its energy producing mitochondria.
A cell with its energy producing mitochondria

Finally, the scientists studied mtDNA, the DNA that comes from cells' energy producers, mitochondria. This DNA is passed directly from women to their children; no mtDNA is inherited from the father. Thus, mtDNA patterns can link people to their mothers and grandmothers and even to ancestors and descendents who are separated by many generations. The mtDNA analysis showed that skeleton 7 was probably the Czarina's. It was related to the skeletons of the three children. In addition, the mtDNA from these four individuals was related to that of Prince Philip, whose grand aunt was the Czarina. (The Prince and the Czarina are related directly and exclusively through the maternal line.) The mtDNA from skeleton 4 (the Czar's skeleton) was like that of two of his living relatives who were directly descended from the Czar's maternal grandmother.

Although the DNA tests indicated that a mother and three children were in the grave, they couldn't prove that the family was the Romanovs. But the location of the grave, the condition of the bones, the finding of gold and platinum fillings in the teeth (only available to the rich), the relations of the DNA samples from the grave to DNA samples from the descendents of the Imperial family, and other evidence all strengthen the case that the bones of the Romanovs had been found.

As for Anastasia, the consensus is that Anna Anderson Manahan was not Anastasia. Two groups of scientists -- one in England and the other in Germany -- tested DNA samples extracted from her preserved blood and intestines and compared them with samples from the bones taken from the Ekaterinburg grave. (Manahan had been cremated, so her bones were not available for testing.) The DNA from the woman who had claimed she was Anastasia did not resemble the DNA of the Czar and Czarina (4). Case closed, at least for Anna Anderson Manahan.

References

    1. Anastasia, The Riddle of Anna Anderson, Peter Kurth (Little, Brown).
    2. Times of London, May 10, 1992, Bahn and Rayment (2 articles).
    3. Nature Genetics 1994, 6:130.
    4. Washington Post, October 6, 1994, B1.


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