Old Dominion University/BIOL294-Genetics/Rinehart-Kim/Romanov Project
Internet Resources
You may use other Internet sources here, but please cite any sources that you use unless they are one of the following.
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0004838 (You should be able to access the entire article. You may need to copy and paste the site address.)
http://www.ncbi.nlm.nih.gov/pubmed/20557352 (You won’t be able to access the entire article, but the abstract will give you important information.)
http://www.nature.com.proxy.lib.odu.edu/ng/journal/v9/n1/pdf/ng0195-9.pdf (Please note that this is a PDF of the article and is provided via the ODU Library.)
History
1. Nicholas II was the last Romanov to hold power in Russia. What was his title? Saint Nicholas the Passion-Bearer, the last Emperor of All Russia, or Tsar Nicholas II, Nicholas the Bloody
2. How long had the Romanov family been in power in Russia? 1613-1917, 304 years
3. Politically, what happened to Nicholas II? His public support collapsed in the wake of WWI and the Russo-Japanese war but was also strongly due to the anti-semitic pogroms, Bloody Sunday, the violent suppression of the 1905 Russian Revolution and the repression of political opponents, the defeat in the Russo-Japanese War which annihilated the Russian Baltic Fleet, and the loss of Russian influence over Manchuria and Korea. He made a lot of wrong moves. He helped start the Great War (now known as World War I) and suffered severe losses, sparking the February Revolution where the Bolsheviks wiped his entire bloodline off the face of the Earth including him shortly after his abdication.
4. Who took control after Nicholas II abdicated the throne? Vladimir Lenin who led the Bolsheviks creating the Soviet Union
5. What happened to Nicholas II and his family after he abdicated the throne? Him and his family were assassinated/executed in Ipatiev House ending the Romanov dynasty
6. One of the reasons that the family of Nicholas II was executed (vs. just imprisoned) was because there was a fear that the White Russian Army would save them. Who was the White Russian Army? Anti-communists who fought the Bolsheviks in the Russian Civil War
Hemophilia
The pedigree chart found at the end of this assignment comes from the Module PowerPoint lecture notes.
7. How was Nicholas II’s wife, Alix, related to Queen Victoria of England? Alix is granddaughter of Queen Victoria, making Nicholas II a “grandson” of Queen Victoria of England
Both Queen Victoria and Alix are designated as being carriers for hemophilia.
8. In a couple of sentences, describe the physiology of the disease hemophilia. (Yes, I know it is severe bleeding because the blood cannot clot. But WHY can’t the blood clot? Be very specific.) Hemophilia changes a gene that provides instructions for clotting in blood, the mutation can eliminate the gene altogether. Bleeding can occur in joints of the body causing swelling that leads to tightness or pain. It can also occur in soft tissue and muscle, mouth, gums, after circumcision, after vaccinations, in urine or stool, and can cause nosebleeds. Hemophilia is an X-linked recessive disorder with female carriers that have males who show symptoms. Coagulation is stopped due to the platelet plug not being stabilized by a fibrin matrix over its surface.
9. What does it mean to be a carrier for a disease? Carriers are capable of passing on genetic mutations of the disease and do not display signs or symptoms of the disease. Carriers are called carriers because they carry or possess the disease in their genetic makeup.
10. What type of hemophilia (A or B) is (probably) represented in the pedigree chart? Hemophilia B
11. On what chromosome is the gene that, when mutated, causes hemophilia? The X chromosome
12. Describe the mutation (at the molecular level) that apparently caused hemophilia in Alix, (and probably all of the European families that had hemophilia). Be very specific. The NCBI article states that a substitution mutation on exon 4, gene F9 was responsible for Alix’s hemophilia. The substitution mutation swaps one nitrogen base for another and this could have changed the amino acid sequence which caused the disease.
13. How could the mutation you described in #12 result in a faulty gene product. Be very specific in your description. This specific mutation is a substitution mutation and could have caused a faulty gene product by changing the amino acid sequence and consequently, the protein produced. It is most likely a substitution of the first or second base of the codon due to the wobble principle. The wobble effect states that a change in the third base pair would likely be silent so that leaves the first and second base pairs.
14. The Romanov’s son, Alexis, had hemophilia. Describe how Alexis genetically acquired hemophilia. (Use a Punnett square. You can either draw a table or line up the genotypes.)
| Romanov Punnett | XH | Y |
| XH | .XHXH | XHY |
| Xh | XHXh | XhY |
Alexis had hemophilia because his mother was a carrier for hemophilia.
15. Using a Punnett square (again, draw a table or line up the genotypes), explain why only males in the pedigree chart have hemophilia. (Choose at least one of the males represented in the pedigree chart and show his parents in the Punnett square.)
| Romanov Punnett | XH | Y |
| XH | XHXH | XHY |
| Xh | XHXh | XhY |
This is the punnett square of Rupert. As you can see males are more likely to have hemophilia if their mother is a carrier. Males have a 50% chance while females would have a smaller chance in most instances.
16. Is it possible for a female to inherit hemophilia, and, if so, how? Yes it is possible for a female to inherit hemophilia, but it is more less likely than males. This is because their mother would have to be a carrier for hemophilia or have the disease, in addition to the father also having hemophilia.
17. Some historians speculate that Alexis’ hemophilia condition could have led to the Russian Revolution. Explain. You should look up the faith healer Rasputin and read about his relationship to the Romanov family.
Tsar Nicholas II sought out a healer for his son Alexis. This healer was named Rasputin and was a faith healer. Rasputin influenced government affairs while attending to Alexis and this influence is what led the public to be weary and could have been a catalyst for eventual revolution.
Molecular Analysis of People in a Mass Grave
18. Two “graves” were discovered near Yekaterinburg, Russia. Describe the number of bodies in each grave. One grave had nine bodies in it and the other had two bodies in it.
19. When were these graves discovered? The first grave was discovered in 1991 while the second was discovered in 2007.
20. What type of testing was done to confirm sex and familial relationships among the remains found in the mass grave? Nuclear DNA testing of five STR markers was performed in order to determine sex and familial relationships among the remains found in the grave.
21. Genetically, what does STR “stand” for? Be very specific in your answer. STR stands for short tandem repeat and is used as a marker during forensic analysis and investigation.
22. Mitochondrial DNA testing was also done on both Nicholas II and Alix. Why was information from Alix’s, but not Nicholas’ mitochondrial DNA used to identify three females as belonging to Alix? The reason Alix’s mitochondrial DNA was used to identify the three females is because mitochondrial DNA is passed on from mother to child, while paternal mitochondrial is not inherited.
23. HRH Prince Philip, the Duke of Edinburgh, provided mitochondrial DNA used to identify Alix and her three daughters. HRH Prince Philip, the Duke of Edinburgh, is married to Queen Elizabeth II of England. Wait, isn’t Queen Elizabeth II related to Queen Victoria? So why was Prince Philip’s mitochondrial DNA used? Based on the pedigree chart, Prince Philip’s DNA was used because he was the grandnephew of Tsarina Alix; being the grandson of one of the many siblings of Alix, specifically an unidentified sister of the line of children of Princess Alice, who is a daughter of Queen Victoria and Edward.
24. Who was missing from the mass grave (the one with the most skeletons)?
Two children were missing from the grave, Alexis and an initially unidentified sister, later theorized to be Anastasia (Coble, M).
Molecular Analysis of People in a Mass Grave, cont.
25. The Duke of Fife and Princess Xenia provided mitochondrial DNA used to identify Nicholas. One of these is a female and another is a male. Does that matter? What general statement can you make about their genetic
relationship to Nicholas and Alexandra? Are these people still living? It does not matter because they are using the mitochondrial DNA in this analysis as the two people they are testing are related to Nicholas (siblings), so would all have the same mitochondrial DNA coming from the mother. Gender would only matter if they were trying to identify Nicholas’s parents.
26. What was discovered in the mitochondrial DNA of Nicholas that was not identified in either the Duke of Fife or Princess Xenia? A point of heteroplasmy at position 16169 (C/T = “Y”) was found in Tsar Nicholas mitochondrial DNA, whereas the Duke of Fife and Princess Xenia’s mitochondrial DNA samples displayed 16169 T (Coble, M).
27. What is the term given to the existence of two (or more) genetically different mitochondria in the cell? When two or more genetically different mitochondria are identified, the condition is referred to as heteroplasmy (Stefano, G).
28. What three types of DNA were used to test the remains found in a second grave? 144.1, 146.1, and 147 (Coble, M).
29. Of the three types of DNA you listed in #28, which one would have been used specifically to identify Alexis? 146.1 (Coble, M.).
30. What was the source of the DNA used to identify Alexis? Two bone fragments from a femoral bone, which was used to identify the 146.1 DNA sample to be male (Coble, M).
31. Was Anastasia in the grave in which Alexis was found? The closest piece of evidence that could confirm that the female bone fragments from the same grave of Alexis was Anastasia would be the other DNA sample of 147 (Coble, M).
Who Wants to Be Anastasia?
Apparently, about 200 people have wanted to be Anastasia and have claimed to be her! One of the most famous imposters was a woman named Anna Anderson (Manahan).
32. Give a brief history (2-3 sentences) of Anna Anderson-both her claims and what is thought to be true. She both claimed and it was speculated that she was Anastasia, and it was also proposed that she was Franzisca Schanzkowska, which was proven true. The mystery carried on as Anastasia’s body was never discovered with the rest of the dead royals, so it was a possibility she was alive. She looked similar to Anastasia which deepened the mystery. Later DNA evidence would prove she was lying and was not Anastasia but was Franzisca Schanzkowska.
33. Where in the US did Anna Anderson eventually settle? Charlottesville, Virginia (Onion, A et al, 2010)
34. Whom did she eventually marry? American history professor, J.E. Manahan (Onion, A et al, 2010)
35. What were the sources of Anna Andersons’s nuclear DNA? A piece of intestines and Hair.
36. What were the sources of Nicholas’ and Alix’s nuclear DNA? The sources of their nuclear DNA came from the bones they exhumed.
37. What type of analysis was done on DNA from Anna Anderson, Nicholas, and Alix? STR, mitochondrial, and nuclear
38. Anna Anderson’s mitochondrial DNA was compared to the mitochondrial DNA of what two people? Duke of Edinburgh and Carl Maucher
39. A hypervariable region of the mitochondrial DNA was analyzed. Define a hypervariable region? Area where base pairs of nucleotides repeat. subject to many changes (Hypervariable region, 2001).
40. What were the conclusions from the mitochondrial DNA comparisons? Anna was not related to the Romanovs, but they did find her true identity as Franciscan Schanzkowska. It was also discovered that she was related to Carl Maucher.
41. The article which describes the analysis of Anna Anderson’s DNA was published in 1995.
When were all of Nicholas’ and Alix’s children finally accounted for? The final bodies were discovered in 2007.
42. What did you learn from doing this assignment? (Each person in a group should answer this question. It is not a group answer.)
Nicole- I learned how we can use DNA testing to solve mysteries. It helps solve problems when people have amnesia or are lying, and you can trace them through relatives and the greater public.
Joshua – I have come to learn how even such small fragments of decades old remains can still be traced to their origins, and in the case of the archaeological find, it is of tracing the very family unit of the Romanovs of the Russian monarchy. I have also learned how such crucial and vital genetic information can still be salvaged from what remained of the Romanovs such as the mitochondrial DNA samples that were successfully recovered but were still prone to scrutiny as to identify which of the Romanovs were buried.
John – What I found most interesting about this assignment was the use of DNA testing. Specifically, the use of Alix’s mitochondrial DNA to identify the bodies. The applications of mitochondrial DNA are very interesting.
Ben- This assignment gave me a history of the royal family and it’s connections, I did not know that hemophilia was carried in it or that the Russian family was connected to it. The use of science to uncover these age-old mysteries and to uncover the past’s mysteries using modern technology is astounding.
Are you still interested in the life of the last Tsar of Russia and his relationship to British royalty? The headline for the following article showed up on my Internet browser earlier this year. While I can’t vouch for it as it did not appear in a peer-reviewed journal, it might be interesting reading for you.
WHILE THIS IS A GROUP PROJECT, EACH STUDENT SHOULD UNDERSTAND EACH QUESTION AND ANSWER GIVEN. BY PLACING YOUR NAME BELOW (AND INCLUDING A DATE), YOU HAVE REVIEWED THE ANSWERS ON THIS ASSIGNMENT, AND AGREE TO THEM.
Nicole Feaster 4/26/2021
Joshua Calderon 4/26/2021
Ben Gustafson 4/27/2021
John Gresham 4/27/2021
REFERENCES
Coble, M., Loreille O., Wadhams, M., Edson, S., Maynard, K., “Mystery Solved: The Identification of the Two
Missing Romanov Children Using DNA Analysis.” Plos One, (2009).https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0004838
Fuhrmann, J. T. The complete wartime correspondence of Tsar Nicholas II and the Empress
Alexandra: April 1914-March 1917. Choice Reviews Online 37, (1999).
Hypervariable Region. Encyclopedia of Genetics 991 (2001). doi:10.1006/rwgn.2001.1868
Onion , A. & Sullivan, M. Woman claiming to be Anastasia Romanov arrives in the U.S. (2010). Available
at: https://www.history.com/this-day-in-history/anastasia-arrives-in-the-united-states. (Accessed: 24th April 2021)
Stefano, G., Bjenning, C., Wang, F., Wang, N., et al. “Mitochondrial Heteroplasmy.” Mitochondrial Dynamics in
Cardiovascular Medicine, Advances in Experimental Medicine and Biology, vol 982. Springer Link, (2017). https://link.springer.com/chapter/10.1007%2F978-3-319-55330-6_30
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