Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 106 (13), 5258-63

Genomic Identification in the Historical Case of the Nicholas II Royal Family

Affiliations

Genomic Identification in the Historical Case of the Nicholas II Royal Family

Evgeny I Rogaev et al. Proc Natl Acad Sci U S A.

Abstract

Accurate unambiguous identification of ancient or historical specimens can potentially be achieved by DNA analysis. The controversy surrounding the fate of the last Russian Emperor, Nicholas II, and his family has persisted, in part, because the bodies of 2 children, Prince Alexei and 1 of his sisters, have not been found. A grave discovered in 1991 contained remains putatively identified as those of the Russian Royal family. However, not all family members were represented. Here, we report the results of genomic analyses of new specimens, the human remains of 2 burned skeletons exhumed from a grave discovered in July 2007, and the results of a comprehensive genomic analysis of remains from the 1991 discovery. Additionally, approximately 117 years old archival blood specimens from Nicholas II were obtained and genotyped, which provided critical material for the specific determination of individual identities and kinship identifications. Results of genotypic analyses of damaged historical specimens were evaluated alongside samples from descendants of both paternal and maternal lineages of the European Royal families, and the results conclusively demonstrate that the recently found remains belong to children of Nicholas II: Prince Alexei and his sister. The results of our studies provide unequivocal evidence that the remains of Nicholas II and his entire family, including all 5 children, have been identified. We demonstrate that convergent analysis of complete mitochondrial genome sequences combined with nuclear DNA profiles is an efficient and conclusive method for individual and kinship identification of specimens obtained from old historic relics.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Complete mt genome sequences retrieved from the putative remains of Prince Alexei (N146), his sister (N147) (second grave) and their parents, Empress Alexandra (N7) and Emperor Nicholas II (N4) (first grave). Short amplification products are shown inside of the mt genome circle. The complete mtDNA sequences from N7, N146 and N147 specimens were identical and each is 2 nt longer than the mtDNA sequence from the N4 skeleton specimen because of insertion of 2 nt in positions 524.1 (A) and 524.2 (C).
Fig. 2.
Fig. 2.
Analysis of maternal lineages of Romanov family. Pedigree represents 2 maternal lineages of Romanov family. The mt haplotype of Nicholas II Romanov is inherited from Empress Maria Feodorovna (Princess Dagmar, the daughter of Louise of Hesse-Cassel and Christian IX, King of Denmark). The haplotype of Empress Alexandra Feodorovna is inherited from Princess Alice, the daughter of Queen Victoria. The complete mitochondrial genome sequences were retrieved from the bone fragments from the second grave: N146, putative Prince (Tsarevich) Alexei, and N147, putative Grand Duchess Maria; and from the first grave: skeleton N7, putative Empress (Tsarina) Alexandra Feodorovna, and skeleton N4, putative Emperor (Tsar) Nicholas II. Complete mt genome sequences or HVR sequences were also determined for the maternal relatives. Large arrows denote individuals with complete mtDNA sequences and small arrows show individuals with HVR sequences determined in this study.
Fig. 3.
Fig. 3.
Analysis of paternal lineages of Romanov family. Y profiles were determined for putative remains of Alexei and Nicholas II and their living cousins of unbroken male lineages of Nicholas I (shown in red) (SI Materials and Methods).
Fig. 4.
Fig. 4.
MtDNA analysis of Nicholas II bloodstain specimen and specimen of skeleton N4. (A) the shirt of Nicholas II with visible traces of bloodstains used for DNA extraction. (B) Comparison of sequence chromatograms of Nicholas II archival sample and 4-46 femur; heteroplasmy was found in both bone and blood specimens.

Similar articles

See all similar articles

Cited by 8 articles

See all "Cited by" articles

LinkOut - more resources

Feedback