The Romanovs

by Robert K. Massie

  Dr. Gill, the head of Biological Services (Research) of the Forensic Science Service, is a slightly built man in his early forties, about five feet nine, with a pale face, uncombed hair, a brown mustache, and watchful eyes behind thick spectacles. He owns a dark blue suit, which he wears for press conferences, but in and around his laboratory his typical dress is a tattered sweater, shapeless corduroys, and elderly loafers. Born in Essex, Gill did an undergraduate degree in zoology at Bristol University, received a doctorate in genetics from Liverpool University, and did a five-year postdoctoral fellowship in genetics at Nottingham University. In 1982, he joined the Forensic Science Service Research Laboratory at Aldermaston to work on forensic applications of conventional blood typing methods. In 1985, against strong opposition in the service, he began to study the utility of DNA profiling in forensic science. Aware of the significance of Alec Jeffreys’s work, he briefly joined Jeffreys’s laboratory and, that same year, coauthored with Jeffreys the first scientific paper which demonstrated that DNA profiling could be used in forensic science. The methods described in this paper are now routinely used around the world. Gill himself has published over seventy papers in the scientific literature.

  Although he is shy and speaks cautiously with strangers, there is one point on which Dr. Gill is quietly emphatic: his laboratory is the best of its kind in the world—“We have retained our world lead” is his way of putting it. In his opinion, therefore, it was entirely understandable that Pavel Ivanov had wished to bring the Russian bones to Aldermaston. “Ivanov asked me a long time ago whether we’d be interested in carrying out these tests,” Gill said. “When he asked, I had to go through the Home Office. They considered all the political ramifications, and eventually we got the go-ahead.”

  The political ramifications existed on many levels. The most obvious was the current relationship between John Major’s Conservative government in Britain and Boris Yeltsin’s presidency of Russia. Both parties were interested in bringing to fruition a long-suspended diplomatic project: a visit to Russia by the queen. No British monarch had visited Russia since 1908, when King Edward VII and Queen Alexandra came by yacht to Tallinn (then Reval) to visit Tsar Nicholas II and Empress Alexandra.* Mikhail Gorbachev and Boris Yeltsin both had invited the queen to come, and Her Majesty and the British Foreign Office wanted the visit to take place.

  But first there was some unfinished historical and family business. The Russian Imperial family and the British Royal family were closely related. King George V, Elizabeth II’s grandfather, was Nicholas II’s first cousin. Indeed, so close was the physical resemblance between the cousins that at George’s wedding, Nicholas often was mistaken for the groom. King George also was a first cousin of Empress Alexandra. In the spring of 1917, after the tsar had abdicated and while Alexander Kerensky and the Provisional Russian government were trying to provide for the safety of the Imperial family by sending them to political asylum abroad, King George V at first welcomed a proposal that Britain bring his Russian cousins to safety by ship. Then the king—fearing that the former tsar’s unpopularity in Britain would tarnish the British monarchy—reversed himself and insisted that they not be brought. George V’s act helped doom Nicholas, his wife, and his five children. When the British door slammed shut, Kerensky sent the family to Siberia, hoping to put them out of reach of the Bolsheviks. They still were there when, seven months after Kerensky’s fall, Lenin’s long arm reached out.

  This catastrophe led to many recriminations. Members of the Russian Imperial family who escaped, aristocratic emigres, and numerous White Russians abroad bitterly condemned King George and his family and descendants. For three quarters of a century, many Russians have regarded England with deep suspicion and resentment. The British Royal family is aware of this hostility. Over the years, palace officials have attempted to bury the king’s role in the Romanov tragedy; official biographers of George V were advised that they should “omit things and incidents which were discreditable.” In 1992, the possibility that the Romanov bones might come to Britain to be verified by British scientists with the help of British Royal persons offered an opportunity to put some of these passionate feelings to rest.

  According to a Forensic Science Service spokesperson who stays close to Dr. Gill specifically to answer nonscientific questions, the decision to bring the bones to Aldermaston was made on a relatively low level; that is, by Janet Thompson, the FSS director general. “Of course,” said the spokesperson, “with the high profile that came with this project, we put it before the home secretary. He could have objected if he had wanted to.” The spokesperson does not know whether Kenneth Clark discussed the project with the foreign secretary or the prime minister. Or whether anyone thought to consult the Royal family. If this was not done, however, Dr. Thompson and Secretary Clark were assuming historical and diplomatic responsibilities far beyond the normal range of their professional and political assignments.

  There was one area in which Thompson—no doubt supported by Clark—did make a decision on her own. This was the decision to ignore the new Thatcherite decree that the FSS was to charge for services and attempt to make a profit. The service spent a large sum of money on the Romanov project. “We did all nine of them, the whole lot,” said Peter Gill. “It managed to be expensive.” “It was very expensive,” chimed in the spokesperson, adding that no figure is available. The sum can be roughly estimated. A year later, the FSS negotiated with a private citizen to perform DNA testing on an unknown woman and a possible relative. These tests were to be performed on preserved tissue and recently drawn blood, both sources from which DNA is far easier to obtain than from old, long-buried bones. For this work, the FSS demanded a five-thousand-pound down payment, plus another five thousand pounds placed in escrow in an English bank. All of this money was spent. The Romanov project involved typing and comparing bone fragments from nine people in Russia, plus blood samples from at least three relatives alive today. Even using the same expense figures for much more difficult tests, this would mean that twelve DNA profiles would cost sixty thousand pounds (over $100,000). Dr. Alka Mansukhani, an American molecular biologist routinely doing DNA extraction and sequencing at New York University Medical Center, believes that, if overhead was included, the figure probably is accurate.

  The Home Office and FSS accountants funded these costs as pure research.

  An adult human body is a cohesive mass of 80 trillion cells, yet in all this amplitude and diversity, there is an extraordinary sameness: each one of these cells contains all the genetic information needed to produce a complete and unique human being. This hereditary knowledge is carried in the chromosomes; in a normal person, forty-six in each cell nucleus, twenty-three from the mother, twenty-three from the father. Chromosomes are made up of molecules of DNA (deoxyribonucleic acid), which use their own chemical structure to store genetic information and commands. The DNA molecules are created from four basic chemical building blocks called bases, and the sequence in which these bases occur provides the information necessary to commence and control the building of a human body. For simplicity’s sake, molecular biologists describe the four bases by their initial letters, A, G, C, and T (adenine, guanine, cytosine, and thymine). The bases appear in pairs bonded with hydrogen; A bonds with T; G bonds with C; these combinations are known as base pairs. In 1953, James Watson and Francis Crick discovered the detailed, overall molecular structure of DNA. They found long, tightly coiled strands, each resembling a ladder twisted into the form of a spiral staircase. The A, C, G, and T base pairs formed the rungs; the sides of the ladder, to which the rungs were attached, were made up of alternating molecules of sugar and phosphate. Watson and Crick named their discovery the double helix.

  The unique structure of every individual human body is dictated by the different combinations of these four letters in base pairs in the DNA. For example, at some point in the strand, one individual will read A, C, G, T, C, C, T. Another person, in the same part of the strand, will display a different s
equence, say A, T, T, C, A, G, C. Whatever the base pair sequence, each cell in a human body contains the same DNA sequence, storing the same information and commands. But, to avoid massive confusion, nature activates only that part of the command system necessary for the function of that particular cell.

  Each cell with its set of forty-six chromosomes contains approximately 3.3 billion DNA base pairs, strung together in clusters of spiraling double helixes. If one magnified this structure to a humanly visible five characters (A, G, T, C, T) per half inch, it would require a strip of paper 162 miles long to write the entire base sequence of a single chromosome. Approximately 99.9 percent of the 3.3 billion base pairs found in a single cell appear in the same sequence in all human beings; they ensure that all humans possess similar characteristics: two eyes, two ears, one nose, ten toes, blood, saliva, stomach acid, and so on. However, in the remaining 0.1 percent (that is, 3.3 million base pairs), the sequence of these base pairs differs from one person to another. It is the fact that individuals vary at this basic molecular level that now permits scientists to determine which human being was the source of this or that sample of bone or tissue, blood, semen, or saliva.

  In the early 1980s, Dr. Alec Jeffreys, working at Leicester University, first recognized the enormous potential of the DNA variable in human beings to resolve questions of identity. He identified regions within hypervariable areas and used radioactive isotopes, called probes, to create an image on film of the DNA strands extracted from individuals. These visible symbols appear strikingly similar to the bar codes which are printed on packages and cans at every supermarket. The DNA patterns—Jeffreys called them “DNA fingerprints”—could then be used to compare one person’s DNA with that of another. Because children would derive half of their DNA base pairs from their mother and half from their father, family relationships could be established or refuted. In 1983, a boy was refused entry into England because an immigration officer doubted that he was the son of a Ghanaian woman who had rights to settlement in the United Kingdom. Jeffreys’s new DNA technique was employed and proved that the boy was the woman’s son. The chance of this match occurring at random was one in ten million.

  Within less than a decade, DNA typing has become the most powerful forensic science tool since the nineteenth-century discovery that the fingerprints of no two persons are the same. Comparisons of DNA now routinely solve paternity cases. Murderers are identified by samples of blood, hair, other tissues, or fluids, liquid or dried. Samples of DNA from bones and teeth have helped resolve long-standing mysteries involving missing persons and unidentified bodies. DNA is remarkably stable: it has been extracted and identified from a three-thousand-year-old Egyptian mummy, from a seven-thousand-year-old mammoth, from the dried saliva remaining on a licked postage stamp. And, properly handled and identified, it is unerring. No prosecutor or defense attorney, no historian, no churchman of any faith, no believer in any political ideology, can disprove the essential message of DNA: that every human being is distinct from every other. DNA evidence, declared one American district attorney, is “like the finger of God pointing at someone and saying, ‘You are the one!’ ”

  Because of the age and deteriorated condition of the Romanov bones, Dr. Gill and Dr. Ivanov faced a task radically more difficult than in any previous DNA typing examination. In a sterile environment, they began by grinding away one millimeter of the contaminated outer surfaces of the bones with sand wheels attached to a high-speed electric drill. The remaining bone was frozen in liquid nitrogen, then pulverized to a fine powder and dissolved in various solutions, then centrifuged to release a microscopic quantity of DNA. So paltry and degraded, in fact, were the sample yields that Gill and Ivanov applied an even more recently developed technique called PCR (polymerase chain reaction), in which selected relevant sections of base pair strands are chemically duplicated over and over in a test tube to provide sufficient quantities of DNA material for scientists to study.

  Using nuclear DNA, the Aldermaston team first turned to determining the sex of each of the skeletons. A gene on the X chromosome (females have two X’s) is six base pairs longer than the similar gene on the Y (males have one X and one Y) chromosome. Using PCR, the scientists could obtain sufficient material to measure and determine this six-base-pair difference. The result was a confirmation of the anthropological findings of Abramov and Maples: there were four males and five females. Next, still using nuclear DNA and studying base pair sequences, Gill and Ivanov tested all nine for a family relationship. Short tandem repeat (called STR.) sequences are natural base pair repetitions in certain hypervariable regions of a chromosome—say, T, A, T, T—occurring over and over again. Within a family, these sequences and the number of repetitions tend to be constant; a different sequence or different number of repetitions in each individual sample would indicate that no family group was present. Again, the results were what was to be expected if the bones had come from the Imperial party. In Gill’s words: “Skeletons 3 through 7 exhibited patterns which would be expected in a family group where 4 and 7 were the parents of children 3, 5, and 6.” The other four adults were excluded as possible parents. Further, Gill’s report continued, “If these remains are the Romanovs then … test data indicated that one of the daughters and the Tsarevich Alexis were missing from the grave.” Other tests established paternity. STR DNA patterns from Body No. 4 were found in No. 3, No. 5, and No. 6; thus, the adult male presumed to be Nicholas was confirmed to be the father of the three young women. This was as far as Gill and Ivanov could go using the small quantity of degraded nuclear DNA available. They had established a party of four males and five females. They had established a family: a father, a mother, their three daughters. But to identify these men and women—to give them names—they had to try another tack.

  Fortunately, a second form of DNA is available in human cells. Called mitochondrial DNA, it appears plentifully in units outside the nucleus which function as power stations for the cell. Mitochondrial DNA is inherited independently of nuclear DNA, and whereas nuclear DNA is inherited half from the mother, half from the father, mitochondrial DNA is inherited exclusively from the mother. From mother to daughter, it is transmitted intact, “passing from generation to generation unchanging, like a time machine,” said Gill. “The same genetic code would be shared by mother, grandmother, great-grandmother, great-great-grandmother, and so on.” At all points in this chain, sons possess mitochondrial DNA received from their mothers, but sons cannot pass this mitochondrial DNA along to their daughters or sons. Thus, as a tool for establishing identity, mitochondrial DNA can be used to identify a woman anywhere in a vertical chain of women descended from one another. And it can identify a son of one of these women. But it cannot continue through the male line; with sons the chain is broken.

  Gill and Ivanov extracted mitochondrial DNA from the nine bone samples brought from Russia. The extracts were amplified to workable quantities by using PCR. To their delight, the quality of the DNA sequences obtained, said Gill, was “generally comparable to that produced from fresh blood samples.” Focusing on two different stretches of the DNA sequence normally hypervariable between different humans, and deriving between 634 and 782 base pair letters for each of the nine subjects, the scientists achieved DNA profiles for all nine of the bone samples they possessed.

  Next, they needed contemporary DNA to make comparisons. The search for living relatives began. People at the FSS and the Home Office drew books from libraries and pored over genealogical trees. Someone drew up a list of names of people who would be scientifically suitable and might be approached. In the case of the Empress Alexandra, finding a genetically useful living relative was easy. Alexandra’s older sister, Princess Victoria of Battenberg, had a daughter, who became Princess Alice of Greece. Princess Alice, in turn, produced four daughters and a son. In 1993, only one of these daughters, Princess Sophie of Hanover, was living. The son was Prince Philip, who became Duke of Edinburgh and the consort of Queen Elizabeth II of England. Prince Philip
, Empress Alexandra’s grandnephew, was perfectly suited for a mitochondrial DNA comparison with bone material of the murdered Russian empress. Accordingly, Dr. Thompson, director of the FSS, wrote to Buckingham Palace and asked whether the prince would be willing to help. Philip agreed, and a test tube filled with his blood soon made its way to Aldermaston. The testing was done in those parts of the mitochondrial DNA sequence where the greatest variety between family groups occurs. By November, Gill and Ivanov had results: the match was perfect; the sequence of DNA base pairs between the mother, the three young women, and Prince Philip was identical. Gill and Ivanov knew that they had located the remains of Alexandra Feodorovna and three of her four daughters.

  Confirming the presence of Tsar Nicholas II was far more difficult. The search for DNA material to compare with that extracted from the femur of Body No. 4 was widespread, prolonged, and, in several instances, controversial. The first attempts were made by Pavel Ivanov. It occurred to him that Nicholas II’s younger brother Grand Duke George, who died in 1899 of tuberculosis at the age of twenty-eight, was buried in the mausoleum of the Romanovs, the Cathedral of St. Peter and St. Paul in St. Petersburg. Comparison of DNA between brothers would nicely suffice. From England, Ivanov contacted Anatoly Sobchak, the mayor of St. Petersburg, and Vladimir Soloviev, who would become the investigator assigned to the Romanov case. “They protested that it would be too expensive,” Ivanov recalled. “ ‘The tombs in the fortress are made of Italian marble.… You must break it.… Who will pay for this?’ And so on.” For eight months, Ivanov persisted, and, at one point, Mstislav Rostropovich, the cellist and conductor, who is a friend of Sobchak, seemed about to pay for the exhumation of Grand Duke George.