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
an article.)

History
1. Nicholas II was the last Romanov to hold power in Russia. What was his title? Nicholas II died as the
“Tsar Nicholas II”
2. How long had the Romanov family been in power in Russia? The Romanov family had been in power
from 1613 to 1917. They ruled for three centuries.
3. Nicholas II abdicated the throne. Who took power then? Nicholas II attempted to give his brother the
throne but his brother said no and did not accept it. Eventually, Lenin took over and created the communist
Russian party.
4. What happened to Nicholas II and his family after he abdicated the throne? After Nicholas II abdicated
his throne him and his family were taken to the Ural Mountains and placed under house arrest.
5. 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? The White
Russian Army were known as anti-Bolshevik forces that fought against communists.

Hemophilia
One of the pedigree charts found at the end of this assignment comes from the Module powerpoint lecture
notes.
6. How was Alix, the wife of Nicholas II, related to Queen Victoria of England? (Look at the pedigree
chart carefully.) The wife of Nicholas II is related to Queen Victoria because that is her grandma.
7. On what chromosome is the gene that, when mutated, causes hemophilia and how does this contribute to
its inheritance pattern? The X chromosome is the gene, when mutated, causes hemophilia.
Both Queen Victoria and Alix are designated as being carriers for hemophilia.
8. What does it mean to be a carrier for a disease? A carrier is an individual who carries and can pass on a
genetic mutation associated with a disease and may or may not display disease symptoms. Carriers are
associated with diseases inherited as recessive traits.
https://www.genome.gov/genetics-glossary/Carrier
9. Why aren’t males considered carriers for hemophilia? There are no genes for clotting factors on the Y
chromosome. This means that males only have one allele for factor VIII and one allele for factor IX. Thus, if a
male has a hemophilia allele on his only X chromosome, he will have the disorder. Females have two X
chromosomes.
https://www.cdc.gov/ncbddd/hemophilia/inheritance-pattern.html
10. 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, in
short, is a disease that inhibits the body of normal blood clotting. This inability for blood to clot in hemophiliacs
is due to an absence of blood clotting factors eight or nine, depending on if you have hemophilia A or B
respectively. Hemophilia is an x-linked genetic mutation and is hereditary in nature.” CDC
https://www.cdc.gov/ncbddd/hemophilia/facts.html
11. What type of hemophilia (A or B) is (probably) represented in the pedigree chart? I believe Hemophilia
B is most likely present in the pedigree chart.
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. According to the NCBI article this mutation
that caused hemophilia in Alix was known as a substitution mutation on exon 4, gene F9. A substitution
mutation works in such a way that one nitrogenous base is swapped for a different one. This substitution
mutation must have caused a change in 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. For this type of mutation to cause faulty product would mean it would have to code for a different
amino acid, thus a different protein. It is most likely that this base was not the third one in the codon because,
thanks to the wobble principle, a change in the third base pair would most likely result in a silent mutation.

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.) The reason that Alexis has
hemophilia is because his mother was a carrier.
Romanov Punnett Square X^H Y
X^H X^H X^H X^H Y
X^h X^H X^h X^h Y

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.) The reason males have hemophilia is because in the Punnett Square it
shows that males are most likely to get it from their mothers who are carriers.
Romanov Punnett Square X^H Y
X^H X^H X^H X^H Y
X^h X^H X^h X^h Y

16. Is it possible for a female to inherit hemophilia, and, if so, how? Yes, it is very possible for a female to
inherit hemophilia. In order for this to be possible the mom needs to be a carrier or has hemophilia herself as
well as the dad would need to have hemophilia also.
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 had a sick son, Alexis. His son needed healing, so the family looked for help
from the Rasputin. Rasputin is known to be a faith healer. Rasputin used his connections to influence many
governmental affairs. Some historians allude to the idea that Rasputin used his connections to favor his self-
interest. This reportedly made the public weary and question those in power and could have been a catalyst to
the 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. There were two graves the first one had nine bodies in it and the second one only had two bodies in it.
19. When were these graves discovered? The grave with 9 people was found in 1991 and the grave with
only two people was found 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 were conducted in order to confirm not only the sex
but also the family relationships.
21. Genetically, what does STR “stand” for? Be very specific in your answer. STR is an acronym for short
tandem repeat and is known as a marker in order to do forensic analysis.
22. 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? (To help you answer this question, look at the second pedigree chart.) Prince Philip’s mitochondrial
DNA was used because there was a maternal relationship, in the mitochondrial DNA, between him and Alix
that was discovered.

23. Who was missing from the mass grave (the one with the most skeletons)? Two children of Nicholas II
and Alexandra are missing; they are Alexis and one of his sisters Maria/Anastasia

Molecular Analysis of People in a Mass Grave, cont.

24. 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 whether it is
male or female, since both have the common ancestor, Louise of Hesse Castle. All of them share the
mitochondrial DNA originated from Louise of Hesse Castle, and hence, have the same mitochondrial DNA.
25. What was discovered in the mitochondrial DNA of Nicholas that was not identified in either the Duke of
Fife or Princess Xenia? They found a single point heteroplasmy in the mtDNA of Nicholas.
26. What is the term given to the existence of two (or more) genetically different mitochondria in the cell?
Heteroplasmy
27. What three types of DNA were used to test the remains found in a second grave? The three types of
DNA used to test the remains in the second grave were: mitochondrial DNA testing, Autosomal STR testing,
and Y-STR testing.
28. Of the three types of DNA you listed in #28, which one would have been used specifically to identify
Alexis? They used Y-STR testing and had an exact match to relatives.
29. What was the source of the DNA used to identify Alexis? A femur fragment.
30. Was Anastasia in the grave in which Alexis was found? It is honestly unknown if Anastasia was in the
grave it is either her or Maria they only know it is a female sister.

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).
31. Give a brief history (2-3 sentences) of Anna Anderson-both her claims and what is thought to be true.
Anna Anderson is a liar who pretended to be Anastasia which was one of her biggest claims. She was actually
born in Poland and her real real name is Franziska Schanzkowska .
32. Where in the US did Anna Anderson eventually settle? Anna Anderson settled down in Charlottesville,
Virginia which is surprisingly only a couple hours away from ODU.
33. What were the sources of Anna Andersons’s nuclear DNA? The sources of Anna Andersons’s nuclear
DNA was samples of hair as well as intestines.
34. What were the sources of Nicholas’ and Alix’s nuclear DNA? The sources for Nicholas’ and Alix’s
nuclear DNA was skeletal bone samples.
35. What type of analysis was done on DNA from Anna Anderson, Nicholas, and Alix? The analysis that
was done is called mtDNA and STR analysis and those were done on all three of them.
36. Anna Anderson’s mitochondrial DNA was compared to the mitochondrial DNA of what two “other”
people? Anna Anderson’s mitochondrial DNA was compared to those of Carl Maucher’s and Tsarina.
37. A hypervariable region of the mitochondrial DNA was analyzed. Define a hypervariable region. A
hypervariable region is defined as.
“a region of a GENOME that is made up of a variable number of repeatedsequences and is diagnostic for an ind
ividual.”, according to The Medical Dictionary.
https://medical-dictionary.thefreedictionary.com/hypervariable+region
38. What were the conclusions from the mitochondrial DNA comparisons? The conclusion was that Anna
was not related to Anastasia and she was actually a polish worker.
39. 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? They were all accounted for in
2007.
40. What did you learn from doing this assignment? I honestly learned so much from this assignment from
not only the history of the Romanov family but how different tests were used in order to identify who was
related. I also found out what hemophilia is linked to as well as seeing it through a pedigree. I really enjoyed
digging for the answers!

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.
https://www.townandcountrymag.com/society/tradition/a31028924/windsors-romanovs-relationship-last-
gathering-true-story/
Source for the pedigree chart above: Janet Stein Carter, Biology Instructor at Clermont College, University of
Cincinnati