Mutations in the Beta Globin Gene and their Effects Holda Anagho and Alexandra Coston 1
The Central Dogma Beta Globin
Beta Globin Nucleotide Sequence (444 nt): ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa): MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH
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.
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.
Beta Globin Mutations Hb Hammersmith Hb Redondo Hb Galicia Hb Showa-Yakushiji Hb Vercelli
Hammersmith 6
Hb Hammersmith Codon 43 7
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
Hb Hammersmith Phenylalanine Serine 9
Hammersmith Wild Type ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Mutation ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTCTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 10
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
More Hammersmith! 12
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
Redondo 14
Redondo! Codon 92 15
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
Redondo Histidine Asparagine 17
Redondo WT ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGAACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 18
Redondo Protein Expression WT 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
Redondo 20
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
Galicia 22
GALICIA! - Codons 97 and 98 23
Galicia Position: chr11 p15.5: 5,247,827-5,247,829 Deletion/Substitution polymorphism Histidine and Valine deleted, Leucine added 24
Hb Galicia Histidine Valine AND DELETED!!! 25
Hb Galicia Leucine ADDED!!!
Galicia WT ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGC TGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA 26
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
Galicia 28
Galicia Effects Hemolytic severe anemia Reticulocytosis Heinz bodies present Found in one Spanish patient 29
Hb Showa-Yakushiji 30
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.
Hb Showa-Yakushiji Codon 110 32
Hb Showa-Yakushiji Proline Leucine 33
Wild Type Hb Nucleotide Sequence (444 nt): ATGGTGCATCTGACTCCTGAGGAGAAGTCTGCCGTTACTGCCCTGTGGGGCAAGGTGAACGTGGATGAAG TTGGTGGTGAGGCCCTGGGCAGGCTGCTGGTGGTCTACCCTTGGACCCAGAGGTTCTTTGAGTCCTTTGG GGATCTGTCCACTCCTGATGCTGTTATGGGCAACCCTAAGGTGAAGGCTCATGGCAAGAAAGTGCTCGGT GCCTTTAGTGATGGCCTGGCTCACCTGGACAACCTCAAGGGCACCTTTGCCACACTGAGTGAGCTGCACT GTGACAAGCTGCACGTGGATCCTGAGAACTTCAGGCTCCTGGGCAACGTGCTGGTCTGTGTGCTGGCCCA TCACTTTGGCAAAGAATTCACCCCACCAGTGCAGGCTGCCTATCAGAAAGTGGTGGCTGGTGTGGCTAAT GCCCTGGCCCACAAGTATCACTAA Translation (147 aa): MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLG AFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEFTPPVQAAYQKVVAGVAN ALAHKYH
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
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.
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
Hb Vercelli 38
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.
Hb Vercelli Codon 126 40
Hb Vercelli DELETED!!! 41
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
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.
Hb Vercelli
Hb Vercelli Effects Symptoms – Mild anemia Gene was found in Italian woman