Part 2

1. Part 2

2. Haemophilia A • An X-linked recessive disorder in which blood clotting does not occur due to deficiency of clotting factor VIII. • In most cases the mutation is the result of insertion of a large segment, consisting of about 3800 bp, in the coding region of the factor VIII. • This results in total inactivation of the protein. SDK 2012

3. Haemophilia A • Inherited as a sex linked recessive trait with bleeding manifestations only in males. • Genes which control factor VIII and IX production are located on the x chromosome • Affected male marries a normal female: none of sons will be affected, all daughters will be carriers • Female carrier marries normal male: 50% chance sons will be affected and 50% chance daughters will be carriers SDK 2012

4. Deletion • Deletions involve removal of one or more base pairs. • They vary greatly in size from deletion of a single base to deletion of a whole gene. • The clinical effects often depend on the size and location of the deleted part of the gene. CTGGAG CT AG SDK 2012

5. Deletion Mutations Normal DNA: CGA – TGC – ATC Alanine – Threonine – stop Mutated DNA: CGA – TCA- TC Alanine – Serine What has happenedto the DNA? A guanine was deleted, thereby pushing all the bases down a frame. What will happen to the amino acids? This is called a deletion mutation, also a type of frameshift mutation. SDK 2012

6. Muscular Dystrophy Deletions of Dystrophin Gene • Dystrophin is a protein that is an important component of skeletal muscle. • The dystrophin gene is located on the p arm of the X chromosome (Xp21.2). • It is a very large gene spanning 2.5 million bp of genomic DNA and consists of 79 exons coding for a protein of approximately 3600 amino acids (11kb). SDK 2012

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8. SDK 2012

9. Muscular Dystrophy Deletions of Dystrophin Gene • Deletion of the whole or most of the dystrophin gene • Dystrophin may not be produced at all • Or produce in in abnormal forms, • Resulting in Duchenne muscular dystrophy • This is a severe X-linked recessive disorder that affects boys and is transmitted by carrier females. X linked Recessive disorder. • In affected boys there is almost complete lack of dystrophin, muscle weakness beginning in childhood and increasing progressively in severity so that the individual is wheel-chair bound at the age of about 15 years. • Death usually ensues in the early twenties due to respiratory muscle involvement. SDK 2012

10. X linked Recessive disorder SDK 2012

11. Becker Muscular Dystrophy(BMD) • Deletions involving a small non-critical part of the gene result in altered dystrophin. • This causes the clinical condition of Becker muscular dystrophy • in BMD muscle weakness begins in adolescence and is very slowly progressive, and affected individuals may lead an almost normal life. SDK 2012

12. Cystic Fibrosis. • Cystic fibrosis (CF) is a genetic condition that affects many organs in the body: especially the lungs, pancreas and sweat glands. • Cystic fibrosis is caused by a mutation in the Cystic Fibrosis Trans-membrane Regulator (CFTR) gene, that is located on chromosome 7. • This gene Produces a trans-membrane protein that regulates the flow of chloride ions into the cells. • The most common mutation is termed the ∆508 mutation, which is a deletion of a single codon at position number 508 in exon 10 of the CFTR gene. • Homozygotes(both parents need to be the carriers of the defective gene) for a ∆508 mutation have cystic fibrosis disease.

13. Gene That Encodes CFTR • The gene that encodes the CFTR protein is found on the human chromosome 7, on the long arm at position q31.2. • Mutations consist of replacements, duplications, deletions or shortenings in the CFTR gene. • This may result in proteins that may not function, work less effectively, are more quickly degraded, or are present in inadequate numbers SDK 2012

14. Cystic Fibrosis. • The defective gene produces a defective protein leading to a blockage in the transportation of the salt, thus leading to production of thick, sticky mucus. SDK 2012

15. Presentation • The secretion of very thick, sticky mucus causes • Obstruction of the bronchi and predisposing to pulmonary infections, • Pancreatic duct obstruction leads to problems with digestion. • Intestinal and liver problems and • When it blocks the sweat glands, it leads to loss of excessive salt through sweat. This leads to imbalance of minerals within the body. SDK 2012

16. Paroxysmal Nocturnal Hemoglobinurea • Paroxysmal nocturnal hemoglobinuria is a disorder of blood cells in which absence of specific molecule(GPI anchor protein, CD55 [Decay Accelerating Factor (DAF)], and CD59 [Membrane Inhibitor of Complement Lysis (MIRL)] on the surface of the cells (particularly RBC) leads to premature destruction of the cells by the complement system. • This destruction is intermittent (paroxysmal). SDK 2012

17. Paroxysmal Nocturnal Hemoglobinurea • GPI anchor protein on the surface of red blood cells produced by the bone marrow stem cells • This is caused by a mutation of PIG-A gene, • PIG-A gene is present on the X chromosome important in making GPI protein anchors. • Defect makes the red cells in susceptible to destruction by the complement system.

18. Paroxysmal Nocturnal Hemoglobinurea • The PIG-A mutation occurs in a bone marrow stem cell. • All the blood cells made by this defective stem cell are deficient in GPI-anchored proteins. (glycosyl-phosphatidylinositol GPI). • The GPI-anchored proteins are present on the surface of red blood cells that protect red cells from the activity of the complement system. • When they are absent , no protection from complement and this lead to intravascular haemolysis.

19. Genetics • PIGA gene(phosphatidylinositol glycan class A) is present in in the X chromosome and can have several mutations, from deletions to point mutations. • The genetic mutation leading to the inability to synthesize the glycosyl-phosphatidylinositol (GPI) anchor proteine.

21. RBC Lysis Normal RBCs PNH RBC CD59 ComplementActivation Chronic Hemolysis Intact RBC Lysed PNH RBCs and free hemoglobin in the plasma

22. Role of CD55 &CD 59

23. Paroxysmal Nocturnal Hemoglobinurea • The term "nocturnal" refers to the belief that hemolysis is triggered by acidosis during sleep and activates complement to hemolyze an unprotected and abnormal RBC membrane. • However, this observation was later disproved. Hemolysis has been shown to occur throughout the day and is not actually paroxysmal, but the urine concentrated overnight produces the dramatic change in color.

24. Deletion of 6 codons in the -globin gene resulting in a variant Hemoglobin • The codons 92 to 97 of the -globin gene are deleted. • This results in a shortened -globin protein that produces a haemoglobin variant termed Haemoglobin Gun Hill. • In homozygotes it produces mild clinical; symptoms. SDK 2012

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26. Frame shift mutations • Frame shift mutations involve a deletion or insertion of one or two base pairs within a coding sequence of a gene. • As the coding message is read in triplets codons and deletions will altered the the reading frame of mRNA • This results in a non-sense sequence of amino acids till stop codon. Original= THE FAT CAT ATE THE WEE RAT Frame-shift= THE FAT CAA TET HEW EER AT SDK 2012

27. Frame Shift Mutations(Deletion) • An example occurs in the -globin gene in which one nucleotide of codon 39 is deleted leads to altered sequence. SDK 2012

28. Frame Shift Mutations(Insertion) • Insertion of a sequence of bases into a coding sequence of a gene. • Sometimes a whole gene sequence may be duplicated. • Hereditary motor and sensory neuropathy type I • A DNA segment at locus 17p11 is duplicated. • Tay-Sachs Disease SDK 2012

29. Tay-Sachs Disease • Tay-sachs disease is an autosomal recessive disorder • This genetic defect is located in the HEXA (hexosaminidase) gene, which is found on chromosome 15. • The hexa gene makes part of an enzyme called beta-hexosaminidase A • This enzyme helps break down a fatty substance called GM2 ganglioside in nerve cells. • Mutations in the HEXA gene disrupt the activity of beta-hexosaminidase A, preventing the breakdown of the fatty substances. • As a result, the fatty substances accumulate to deadly levels in the brain and spinal cord. • The buildup of GM2 ganglioside causes progressive damage to the nerve cells. SDK 2012

30. Tay-Sachs Disease SDK 2012

31. Tay-Sachs Disease SDK 2012

32. Tay-Sachs Disease SDK 2012

33. Tay-Sachs Disease S\S • Loss of hearing • Physical and mental retardation • Seizures • Dementia • And most noticeably detected by the red dots it causes on the retina of an individuals eye SDK 2012

34. Trinucleotide Repeat Expansions • Trinucleotides are triplets of nucleotides that are repeated • The number of repeats varies in different individuals. • Trinucleotide repeats is - - - CAG CAG CAG CAG CAG - - - - • Trinucleotide repeats are widespread in the genome, and may occur in exons, introns, promoter sequences or non-coding regions. • They are perfectly normal and occur in all individuals • However a mutation arises when the repeats become unstable and undergo expansion, namely an increase in the number of repeats as they are transmitted from one generation to the next. • When the number of repeats exceeds a certain limit, clinical symptoms occur. • An example is the huntingtin gene, which, when mutated, causes Huntington's disease. • Normal range of (CAG)n is 11 to 34 • Huntington's disease appear in individuals in whom the number of repeats is greater than 37. SDK 2012

35. Huntington's Disease • Autosomal Dominant Inheritance • Due to an excess of C-A-G nucleotide repeats(>37) in the HTT gene on the short arm of chromosome 4 forms (4p 16.3)Huntington's protein. • Huntington's protein has increase number of glutamine (polyglutamine). • Excess of repeats causes the protein to form aggregates that are deposited within the neurons causing neuronal degeneration • Affects brain and spinal cord, especially the basal ganglia. SDK 2012

36. Trinucleotide Repeat Expansions SDK 2012

37. Clinical Manifestations • Most commonly appear in mid-forties • If appear at a younger age, more severe • Manifestations occur because of wasting away of brain cells. • Sudden jerky, involuntary movements throughout body • Difficulties with balance and coordination • Dysphasia • Hesitant/slurred speech • Progressive dysfunction of intellectual and thought processes (dementia) • Cognitive deficits a. Working memory loss b. Reduced capacity to plan, organize, and sequence 7. Restlessness, irritability 8. Depression or Euphoria SDK 2012

38. What happens to a person who has a mutation? • Most of the time the mutation is harmless because 95% sections of DNA do not code for protein (junk DNA). • But sometimes the mutations can cause disorders such as Huntington’s disease and sickle cell anemia etc. SDK 2012

39. Brain Work 5 Which of these is NOT a type of mutation. a) Point mutation b) Flyaway mutation c) Frameshift mutation d) Nonsense mutation • Flyaway Mutation SDK 2012

40. Brain Work 6 1. THE CAT SAW THE FAT RAT 2. THE CAT SAW THE RAT The change in Statement 1 to form Statement 2 is most similar to what type of mutation? • Insertion • Deletion • Substitution • Frameshift • The correct answer is B deletion. • Because the sentence is missing the word fat which occurs in deletion as it is the removal of a section of DNA. SDK 2012

41. Brain Work 7 5′AGAUCGAGU3′ → 5’ACAUCGAGU3′ The chain above represents three codons. Which of the following changes would be expected in the amino acid chain if the mutation shown above occurred? The amino acid sequence would be shorter than expected. The identity of one amino acid would change. The amino acid sequence would remain unchanged. The identities of more than one amino acid would change. • The correct answer is B because according to the codon chart if G is switched by a C then one amino acid is affected because now instead of coding for Arginine it now codes for Threonine. SDK 2012

42. Brain Work 7 5′AGAUCGAGU3′ → 5’AGAUGAAGU3′ The chain above represents three codons. Which of the following changes would be expected in the amino acid chain if the mutation shown above occurred? The amino acid sequence would be shorter than expected. The identity of one amino acid would change. The amino acid sequence would remain unchanged. The identities of more than one amino acid would change. • The correct answer is A because according to the codon chart if C G are switched by a G A then one codon will be affected because now instead of coding for Serine it now codes for Stop codon. SDK 2012

43. Thank You SDK 2012