1. Mutations in the Beta Globin Gene and their Effects
Holda Anagho and Alexandra Coston 1

2. The Central Dogma Beta Globin

3. Beta Globin
Nucleotide Sequence (444 nt): ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa): MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH

4. Beta Globin Reading Frame
5'3' Frame 1 Y I C F Stop H N C V H Stop Q P Q T D T Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L V S R L Q D R F K E T N R N W A C G D R E D S W V S D R H Stop L S L P I G L F S H P Stop A A G G L P L D P E V L Stop V L W G S V H S Stop C C Y G Q P Stop G E G S W Q E S A R C L Stop Stop W P G S P G Q P Q G H L C H T E Stop A A L Stop Q A A R G S Stop E L Q G E S Met G R L Met F S F P F F S Met V K F Met S Stop E G D K Stop Q G T V Stop N G K Q T N D C I S V E V S G S F Stop F L L F A V H N N C F L L F N S C F L F F S S P Q F L L L Y L Met P Stop H C V Stop Q K E I S L R Y I K Stop L K K K L Y T V C L V H Y Y L E Y Met C A Y L H I H N L P T L F S F I F N Stop Y I I I I H I Y G L K C N V L I C V H I L T K S G Stop F C I C N F K K C F L L L I Y F F V Y L I S N T F P N L F L S G Q Stop Stop Y N V S C L F A P F Stop R I T V I I S G L R Q Stop Q Y L C I Stop I F L H I N C N Stop C K R F H I A N S S Y N P A T I L L L F Y G W D K A G L F Stop V Q A R P F C Stop S C S Y L L S S S H S S W A T C W S V C W P I T L A K N S P H Q C R L P I R K W W L V W L Met P W P T S I T K L A F L L S N F Y Stop R F L C S L S P T T K L G D I Met K G L E H L D S A Stop Stop K T F I F I
5'3' Frame 2 T F A S D T T V F T S N L K Q T P W C I Stop L L R R S L P L L P C G A R Stop T W Met K L V V R P W A G W Y Q G Y K T G L R R P I E T G H V E T E K T L G F L I G T D S L C L L V Y F P T L R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R V S L W D A Stop C F L S P S F L W L S S C H R K G I S N R V Q F R Met G N R R Met I A S V W K S Q D R F S F F Y L L F I T I V F F C L I L A F F F F L L R N F Y Y Y T Stop C L N I V Y N K R K Y L Stop D T L S N L K K N F T Q S A Stop Y I T I W N I C V L I C I F I I S L L Y F L L F L I D T Stop S L Y I F Met G Stop S V Met F Stop Y V Y T Y Stop P N Q G N F A F V I L K N A F F F Stop Y T F L F I L F L I L S L I S F F Q G N N D T Met Y H A S L H H S K E Stop Q Stop Stop F L G Stop G N S N I S A Y K Y F C I Stop I V T D V R G F I L L I A A T I Q L P F C F Y F Met V G I R L D Y S E S K L G P F A N H V H T S Y L P P T A P G Q R A G L C A G P S L W Q R I H P T S A G C L S E S G G W C G Stop C P G P Q V S L S S L S C C P I S I K G S F V P Stop V Q L L N W G I L Stop R A L S I W I L P N K K H L F S L
5'3' Frame 3 H L L L T Q L C S L A T S N R H H G A S D S Stop G E V C R Y C P V G Q G E R G Stop S W W Stop G P G Q V G I K V T R Q V Stop G D Q Stop K L G Met W R Q R R L L G F Stop Stop A L T L S A Y W S I F P P L G C W W S T L G P R G S L S P L G I C P L L Met L L W A T L R Stop R L Met A R K C S V P L V Met A W L T W T T S R A P L P H Stop V S C T V T S C T W I L R T S G Stop V Y G T L D V F F P L L F Y G Stop V H V I G R G Stop V T G Y S L E W E T D E Stop L H Q C G S L R I V L V S F I C C S Stop Q L F S F V Stop F L L S F F F F S A I F T I I L N A L T L C I T K G N I S E I H Stop V T Stop K K T L H S L P S T L L F G I Y V C L F A Y S Stop S P Y F I F F Y F Stop L I H N H Y T Y L W V K V Stop C F N Met C T H I D Q I R V I L H L Stop F Stop K Met L S S F N I L F C L S Y F Stop Y F P Stop S L S F R A I Met I Q C I Met P L C T I L K N N S D N F W V K A I A I S L H I N I S A Y K L Stop L Met Stop E V S Y C Stop Stop Q L Q S S Y H S A F I L W L G Stop G W I I L S P S Stop A L L L I Met F I P L I F L P Q L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop A R F L A V Q F L L K V P L F P K S N Y Stop T G G Y Y E G P Stop A S G F C L I K N I Y F H
3'5' Frame 1 Q Stop K Stop Met F F I R Q N P D A Q G P S Stop Y P P V Stop Stop L D L G N K G T F N R N W T A R K R A Stop Stop Y L W A R A L A T P A T T F Stop Stop A A C T G G V N S L P K Stop W A S T Q T S T L P R S C G R K I R G Met N Met I S K R A Stop L G L R I I Q P Y P N H K I K A E W Stop L D C S C Y Stop Q Y E T S Y I S Y N L Y A E I F I C R D I A I A L T Q K L S L L F F R Met V Q R G Met I H C I I I A L K E R D Stop G K Y Stop K Stop D K Q K S I L K E E S I F Stop N Y K C K I T L I W S I C V H I L K H Y T L T H K Y V Stop Stop L C I N Stop K Stop K K I K Stop G D Y E Y A N K H T Y I P N S N V L G R L C K V F F Stop V T Stop C I S E I F P F V I H N V K A L S I I V K I A E K K K K E S K N Stop T K E N N C Y E Q Q I K E T K T I L R L P H Stop C N H S S V S H S K L Y P V T Y P L P Met T Stop T Stop P Stop K R R G K K T S S V P Stop T H P E V L R I H V Q L V T V Q L T Q C G K G A L E V V Q V S Q A I T K G T E H F L A Met S L H L R V A H N S I R S G Q I P K G L K E P L G P R V D H Q Q P K G G K I D Q Stop A E R V S A Y Q K P K S L L C L H Met P S F Y W S P Stop T C L V T L I P T C P G P H H Q L H P R S P C P T G Q Stop R Q T S P Q E S D A P W C L F E V A S E H S C V R S K C
3'5' Frame 2 N E N K C F L L G R I Q Met L K A L H N I P Q F S S W T Stop G T K E P L I E I G Q Q E S E L S D T C G P G H Stop P H Q P P L S D R Q P A L V G Stop I L C Q S D G P A H R P A R C P G A V G G R Stop E V Stop T Stop L A K G P S L D S E Stop S S L I P T I K Stop K Q N G S W I V A A I S N Met K P L T S V T I Y Met Q K Y L Y A E I L L L P Stop P R N Y H C Y S L E W C K E A Stop Y I V S L L P Stop K K E I R E S I R N K I N K K V Y Stop K K K A F F K I T N A K L P Stop F G Q Y V Y T Y Stop N I T L Stop P I N Met Y N D Y V S I K N K R K Stop S R E I Met N Met Q I S T H I F Q I V Met Y Stop A D C V K F F F K L L N V S Q R Y F L L L Y T Met L R H Stop V Stop Stop Stop K L R R R K K K K A R I K Q K K T I V Met N S K Stop K K L K R S Stop D F H T D A I I R L F P I L N C T L L L I P F L Stop H E L N H R K E G E R K H Q A S H R L T L K F S G S T C S L S Q C S S L S V A K V P L R L S R Stop A R P S L K A P S T F L P Stop A F T L G L P I T A S G V D R S P K D S K N L W V Q G Stop T T S S L R V G K Stop T N R Q R E S V P I R N P R V F S V S T C P V S I G L L K P V L Stop P Stop Y Q P A Q G L T T N F I H V H L A P Q G S N G R L L L R S Q Met H H G V C L R L L V N T V V S E A N V
3'5' Frame 3 Met K I N V F Y Stop A E S R C S R P F I I S P S L V V G L R E Q R N L Stop Stop K L D S K K A S L V I L V G Q G I S H T S H H F L I G S L H W W G E F F A K V Met G Q H T D Q H V A Q E L W E E D K R Y E H D Stop Q K G L A W T Q N N P A L S Q P Stop N K S R Met V A G L Stop L L L A I Stop N L L H Q L Q F I C R N I Y Met Q R Y C Y C L N P E I I T V I L Stop N G A K R H D T L Y H Y C P E R K R L G K V L E I R Stop T K K Y I K R R K H F L K L Q Met Q N Y P D L V N Met C T H I K T L H F N P Stop I C I Met I Met Y Q L K I K E N K V G R L Stop I C K Stop A H I Y S K Stop Stop C T R Q T V Stop S F F L S Y L Met Y L R D I S F C Y T Q C Stop G I K Y N S K N C G E E K K R K Q E L N K R K Q L L Stop T A N K R N Stop N D P E T S T L Met Q S F V C F P F Stop T V P C Y L S P S Y D Met N L T I E K K G K E N I K R P I D S P Stop S S Q D P R A A C H S A A H S V W Q R C P Stop G C P G E P G H H Stop R H R A L S C H E P S P Stop G C P Stop Q H Q E W T D P Q R T Q R T S G S K G R P P A A Stop G W E N R P I G R E S Q C L S E T Q E S S L S P H A Q F L L V S L N L S C N L D T N L P R A S P P T S S T F T L P H R A V T A D F S S G V R C T Met V S V Stop G C Stop Stop T Q L C Q K Q Met
Different reading frames of beta globin gene. The correct reading frame is the 5’3’ Frame 1. There are so many stop codons here because it includes non-coding sequences such as introns and gene regulators.
Theoretically, there are three possible ways an mRNA can be “read.” Each reading frame is capable of producing different amino acids. However, there is only 1 correct reading frame that gets translated.

5. Beta Globin Reading Frame (Cont’d.)
5'3' Frame 1 Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop
5'3' Frame 2 W C I Stop L L R R S L P L L P C G A R Stop T W Met K L V V R P W A G C W W S T L G P R G S L S P L G I C P L L Met L L W A T L R Stop R L Met A R K C S V P L V Met A W L T W T T S R A P L P H Stop V S C T V T S C T W I L R T S G S W A T C W S V C W P I T L A K N S P H Q C R L P I R K W W L V W L Met P W P T S I T
5'3' Frame 3 G A S D S Stop G E V C R Y C P V G Q G E R G Stop S W W Stop G P G Q A A G G L P L D P E V L Stop V L W G S V H S Stop C C Y G Q P Stop G E G S W Q E S A R C L Stop Stop W P G S P G Q P Q G H L C H T E Stop A A L Stop Q A A R G S Stop E L Q A P G Q R A G L C A G P S L W Q R I H P T S A G C L S E S G G W C G Stop C P G P Q V S L
3'5' Frame 1 L V I L V G Q G I S H T S H H F L I G S L H W W G E F F A K V Met G Q H T D Q H V A Q E P E V L R I H V Q L V T V Q L T Q C G K G A L E V V Q V S Q A I T K G T E H F L A Met S L H L R V A H N S I R S G Q I P K G L K E P L G P R V D H Q Q P A Q G L T T N F I H V H L A P Q G S N G R L L L R S Q Met H H
3'5' Frame 2 Stop Stop Y L W A R A L A T P A T T F Stop Stop A A C T G G V N S L P K Stop W A S T Q T S T L P R S L K F S G S T C S L S Q C S S L S V A K V P L R L S R Stop A R P S L K A P S T F L P Stop A F T L G L P I T A S G V D R S P K D S K N L W V Q G Stop T T S S L P R A S P P T S S T F T L P H R A V T A D F S S G V R C T
3'5' Frame 3 S D T C G P G H Stop P H Q P P L S D R Q P A L V G Stop I L C Q S D G P A H R P A R C P G A Stop S S Q D P R A A C H S A A H S V W Q R C P Stop G C P G E P G H H Stop R H R A L S C H E P S P Stop G C P Stop Q H Q E W T D P Q R T Q R T S G S K G R P P A A C P G P H H Q L H P R S P C P T G Q Stop R Q T S P Q E S D A P
This shows the amino acid sequence of beta globin without the noncoding sequences. You can see that in the correct reading frame (5’3’ Frame 1), the sequence starts with Methionine and codes until it reaches a stop codon at the end. There are no interruptions here. Methionine is the start codon, and translation of any mRNA ALWAYS begins with Met. The tRNA scans the mRNA until it finds the first AUG (the codon for Met) and then it starts translating the mRNA.
In the incorrect reading frames, there are many stop codons, so they are obviously not the correct reading frames for the beta globin gene.

6. Beta Globin Mutations Hb Hammersmith Hb Redondo Hb Galicia
Hb Showa-Yakushiji
Hb Vercelli

7. Hammersmith 6

8. Hb Hammersmith
Codon 43 7

9. Hb Hammersmith Hb Chiva Location: chr11 p15: 5,247,994
Single nucleotide polymorphism
Phenylalanine, a nonpolar amino acid, is changed to Serine, an uncharged polar residue. 8

10. Hb Hammersmith
Phenylalanine
Serine 9

11. Hammersmith
Wild Type ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA
Mutation ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTCTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 10

12. Hb Hammersmith
Wild Type Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop
Mutation Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F S E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop 11

13. More Hammersmith! 12

14. Hammersmith Effects Hemolytic severe Anemia Increase in Heinz bodies
Decreased cooperativity
Decreased Oxygen affinity
Found in a few English children, a Japanese male, a North American Indian, a Caucasian woman in Minnesota
Found de novo mutation in black twins 13

15. Redondo 14

16. Redondo!
Codon 92 15

17. Hb Redondo Hb Isehara Location: chr11 p15.5: 5,247,845
Single nucleotide polymorphism
Histidine, a positively charged residue, is replaced with Asparagine, uncharged polar 16

18. Redondo
Histidine
Asparagine 17

19. Redondo WT
ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA
ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGAACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 18

20. Redondo Protein ExpressionWT
Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop
Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L N C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop 19

21. Redondo 20

22. Redondo Effects Unstable beta globin, hemoglobin unstable
Hemolytic chronic Anemia
Reticulocytosis (Increase in immature red blood cells)
Found as a de novo mutation in a Portuguese male and a Japanese female and her daughter 21

23. Galicia 22

24. GALICIA!
- Codons 97 and 98 23

25. Galicia Position: chr11 p15.5: 5,247,827-5,247,829
Deletion/Substitution polymorphism
Histidine and Valine deleted, Leucine added 24

26. Hb Galicia
Histidine
Valine
AND
DELETED!!! 25

27. Hb Galicia
Leucine
ADDED!!!

28. Galicia WT
ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA
ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGC TGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 26

29. Galicia Protein Expression
Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L H V D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop
Met V H L T P E E K S A V T A L W G K V N V D E V G G E A L G R L L V V Y P W T Q R F F E S F G D L S T P D A V Met G N P K V K A H G K K V L G A F S D G L A H L D N L K G T F A T L S E L H C D K L L D P E N F R L L G N V L V C V L A H H F G K E F T P P V Q A A Y Q K V V A G V A N A L A H K Y H Stop WT 27

30. Galicia 28

31. Galicia Effects Hemolytic severe anemia Reticulocytosis
Heinz bodies present
Found in one Spanish patient 29

32. Hb Showa-Yakushiji 30

33. Hb- Showa-Yakushiji Beta+ Thalassemia Location: chr11 5,246,940
Codon 110
Substitution mutation.
CTG CCG
Leucine Proline
Hb Showa-Yakushiji is a Beta plus Thalassemia, which means the beta gene is made but in insufficient quantities
The mutation is located on chromosome 15, nucleotide 5,246,940. On Codon 110, the nucleotide Thymine is changed to Cytosine. This changes the amino acid from Leucine to Proline. CTG (Leucine) is changed to CCG (Proline). Both amino acids are hydrophobic, so this may not drastically change the structure of the protein.

34. Hb Showa-Yakushiji
Codon 110 32

35. Hb Showa-Yakushiji
Proline
Leucine 33

36. Wild Type Hb
Nucleotide Sequence (444 nt):  ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa):  MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH

37. Hb Showa-Yakushiji
Nucleotide Sequence (444 nt):  ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCCGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA
Translation (147 aa):  MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVPVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH
Sequence of Hb-Showa yakushiji. You can see that T changes to C, which changes the amino acid from Leucine (L) to Proline

38. Hb Showa-Yakushiji
Location of mutation on amino acid tertiary structure. Its not close to the heme and it’s hydrophobic like proline, so I would infer that the structure of the protein will not change too much.

39. Hb Showa-Yakushiji Effects
Hemolytic Anemia
Hyperunstable
Low % of Japanese
Low % of United Arab Emirates
Effects; Hemolytic anemia; Anemia due to the abnormal breakdown of red blood cells
My hypothesis is wrong. The change from leucine to proline actually has a drastic effect on beta globin. It makes the structure hyperunstable, which causes the red blood cells to break down prematurely. 37

40. Hb Vercelli 38

41. Hb Vercelli Mutation location codon 126 chr11 5,246,892
Beta 0 Thalassemia
Frameshift mutation
This is a beta0 thalassemia, which means that no beta gene is detected. This mutation is found on codon 126. Nucleotide 5,246,892 on chromosome 11 is deleted. This causes a shift in the reading frame, and changes every amino acid from codon 126 onwards. Also, because the frame is shifted, the stop codon that was located on codon 144 is no longer a stop codon. The mRNA is translated until a new stop codon is found at codon 156. This means that some of the 3’UTR is translated.
A symptom of Hb Vercelli is Mild anemia.

42. Hb Vercelli
Codon 126 40

43. Hb Vercelli
DELETED!!! 41

44. Wild Type Hb
Nucleotide Sequence (444 nt):  ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa):  MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH
The T in red is deleted, which causes a frameshift mutation

45. Hb Vercelli
ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAG GCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa):  MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPGRLPIRKWWLVWMPWPTSITKLAPLLSNP(156)Y-COOH
The T in red is deleted, which causes a shift in frame. This is called a frameshift mutation. All subsequent codons are changed, which means amino acids are changed.

46. Hb Vercelli

47. Hb Vercelli Effects Symptoms – Mild anemia
Gene was found in Italian woman