Gene Mutation

Lecturer : Dr. Ameen Al-Ali :: Done By :: Husam Al-jofi & 3ahad


Insertions & deletions :  A number of inherited diseases are caused by the insertion of many copies of the same triplet of nucleotides .(fragileX, Hungtington's disease). e.g. 100 copies of (AGU) (AGU) (AGU) (AGU) (AGU)…..(triplets)×100 called triplet expansion  Amino acid sequence of the protein subsequent to mutation bears no resemblance to the normal sequence. e.g. Once you enter the glycine code and copy it 200 time the structure of protein will totally change  Most frame shift mutations leads to premature stop codon down stream to the mutation. 90% frame shift → truncation (shortness) of the protein, not complete Frameshift : is that when u add or delete 1,2,4,7…not 3 amino acids therefore it will lead to early stop codon My note # just remember that stop codon is important in DNA translation to prevent elongation of the gene synthesis.

Duplication :  Duplication are doubling of a section of the genome during meiosis. not one base but millions of bases a hole piece of DNA has been duplicated which cause a serious effect  This can cause the gene to carry inappropriate promoters at its 5' end. Promoter : determines how fast the gene is transcribed 1 gene→1 specialized promoter (majority) 1 gene→ with other promoter cause problems Linked genes : 3genes → 1 promoter e.g. 3 enzyms are required to metabolize lactose under the same promoter  Normal cell protein coupled to C-terminal of another protein (leukaemia). take N-protien of a gene and insert it another gene therefore duplication of this new mutated gene will cause leukaemia.


Translocation :  Transfer of a piece of one chromosome to a non-homologous chromosome.  This can alter the phenotype in several ways o Break may occur within a gene destroying its function. if u break the gene in half it will lose its function o Gene may come under influence of different promoters & enhancers. o Breakpoint may occur within a gene creating a hybrid gene. i.e. protein with an N-terminal of one normal cell protein coupled to C-terminal of another (leukaemia). ½ gene + ½ gene of another gene → with totally new function differ then the originals (e.g. leukaemia)

- The gene D will from the long arm to the short arm, and the C gene will move from the short arm to the long 1. and when - A gene in a given chromosome may be controlled by another gene or factor from another chromosome so there is a complicated interaction between genes present on different chromosomes and this what complicates the whole case of genetics


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Single Nucleotide Mutation : Single base substitution (also called point mutation) Missense mutation (sickle cell disease) Non mutation (cystic fibrosis) Silent mutation (one in G6PD deficiency) Due to synonymous(degeneracy of the codon) mutation (is like 1 codon of Arg is replaced by another codon of Arg )& other cause is there is a mutation but it does not led to any phenotypic character or disease e.g. no change in haemoglobin between Hb g (glutamic) & Hb a (alanine), the importance of this is to differentiate between other disease like markers e.g. Hb g are cretin to develop some kind of disease more than Hb a (diagnostic).

- Splice-site mutation (thalassemia) the intron & axon are linked by a splice site that contain (AG) in the beginning & (GA) in the end, and if we have a mutation of the end GA→GT so when the enzyme will continue synthesis till the next intron without stopping cause all he knows is GA.
AG 146 amino acid AG 170 amino acid GA STOP GT Continue with no exon connected till next intron β- thalasemia EXON

- Promoter site (altered expression) No change in protein the change is in the rate of expression (syntheses/transcription) of that protein → effect translation → reduction or increase of the activity of the protein  Insertion & deletion  Duplication(during meiosis)


Point mutation and small deletions : Wild –type sequences N-phe Arg Trp Ile Amino 5’-UUU CGA UGG acid mRNA 3’-AAA GCT ACC DNA 5’-TTT CGA TGG Sense strand is the same as DNA but the AUA


Asp-C AAU-3’

→Small protien

TAT CGG TTA-5’ ATA GCC AAT-3’ difference is T→U

Miessense mRNA 3’-AAT GCT ACC TAT CGG TTA-5’ DNA 5’-TTA CGA TGG ATA GCC AAT-3’ N-Leu Arg Trp Ile Ala Asp-C One of the T is mutated to A the (only change its in the amino acid and no frameshift happened ) which cause sicle cell anaemia Non-sense mRNA 3’-AAA GCT ATC TAT CGG TTA-5’ DNA 5’-TTT CGA TAG ATA GCC AAT-3’ N- Phe Arg Stop TGG →TAG, more over the transfer RNA does not know this codon (TAG) so the protein stops here instead of 6 protiens we have 2 which is known as premature termination (truncation of a protein) e.g. some of β- thalasemia & breast cancer Frameshift by addition mRNA 3’-AAA GCT DNA 5’-TTT N-phe CGA Arg ACC TGG Trp ATA TAT Tyr TCG AGC Ser GTT A5’ CAA T3’ Gln

Frameshift by deletion GCTA CGAT mRNA 3’-AAA CCT DNA 5’-TTT GGA N-Phe Gly



TA-5’ AT-3’


e.g. of missence mutation the only change is that Glu GAG → Val GTG but the rest didn’t change, and by this it will result in forming βSicle cell hemoglobin

e.g of framshift mutation ur read it Glu, Pro, Gln, Leu…etc. but when u delete G in u will read it like this Ser, Arg, Asn, Phe….etc The same thing goes when u add amino acids Or when u add or delete 2 Not 3, 6, 9

Stop codons (termination) : Non of these codons is recognized by tRNA and recognized by protein factors. RF1 ( releasing factor) recognizes UAA & UAG while RF2 recognizes UGA & UAA. they are universal from human to bacteria  UAG : (amber) more error reading this codon . most common in human ,TAG in DNA, amber = yellow  UAA : (ochre) most commonly used termination codon in bacteria ochre = green  UGA : (opal) is the least efficient opal = red


Suppressor mutations: wrong + wrong = right A mutation in the gene for a tRNA molecule that changes its anti-codon loop can "suppress" nonsense mutations that occur elsewhere in protein-coding genes. Generation of a 'Nonsense' Mutation :  The wild type DNA sequence : 5'- CTA CAG ATT -3' 3'- GAT GTC TAA -5' Produces the mRNA 5'- CUA CAG AUU -3' which codes for the poly peptide leu gln iso ..etc. st  A 1 position mutation ( C→ T ) in the second triplet gives a 'mutant' DNA sequence sense strand 5'- CTA TAG ATT -3' 3'- GAT ATC TAA -5' Produces the mRNA which 5'-CUA UAG AUU -3' codes for the polypeptide Leu-* because UAG is a 'stop' codon (the so-called amber stop). Chain growth in the polypeptide terminates prematurely. So all u have is only one codon instead of three which is GAT Generation of an 'amber suppressor' mutation in a tRNA gene :  tDNA sense strand 5'- TAC -3' 3’- ATG – 5’ which produces the tRNA 3'- AUG -5' anticodon loop which reads the mRNA 5'- UAC -3' as –tyrtRNA anticodon AUC combines with mRNA UAC codon and by this we could recognise the UAC mRNA ‫ عهى‬tRNA ‫ع يا فيى كٍف ٌتعزف‬ ‫انهً نس‬ 5'- TAC -3' sense strand ‫بأحاًل أبسط نو انًسأنت‬ 3'- ATG -5' non-sense strand mRNA ‫ انهً بٍتعزف عهى‬antibodies ٌ‫ ى‬tRNA ‫اعتبز‬ 3'- AUG -5' tRNA ‫ انهً جاء ين بزه‬antigens ٌ‫انهً ى‬ 5'- UAC -3' mRNA


 A 3rd-position mutation(C→G) in this region produces a 'mutant'  tDNA gene sense strand 5'-TAG-3' which produces the tRNA 3'-AUC-5' anticodonloop which reads the mRNA a 5'-UAG-3' as -tyrThen, the mutated tDNA gene produces a tRNA molecule that will read the the mutated gln codon UAG as a tyr codon, instead of as a 'stop' codon, this will continue synthesis of the protein without stopping till the next codon.
5'- TAG -3' sense strand 3'- ATC -5' non-sense strand 3'- AUC -5' tRNA 5'- UAG -3' mRNA

Mutation in Non-Coding Region : o Most likely to have a phenotypic effects, expect when it falls in o Promoter region that effect the level of expression o Spilce region o Splice acceptor "AG" , or splice donor "GT" In the picture; IVS 1: intervening sequence 1; i.e. intron 1 If a mutation takes place in an intron ( a non coding region ); it will most likely lead to a phenotypic effect, except if it falls in the promoter site which affects level of expression not structure, while in splice, the number of amino acids will change, which is a structure mutation, i.e. case of thalassemia The beat chain of haemoglobin consists of 146 amino acids, which means that the beta Hb gene consists of 146x3= 450 base approximately Yet, the real size of the beta Hb gene is from 2800 to 3000 bases “nucleotides” Why? Because; 3 exons=450 approximately + an intron which contains further more bases The problem in tahalssemia is a mutation in the splice site leading to an intron between exon 1 and exon 2, which is not supposed to be there, this will lead to coding of more amino acids, around 170 instead of 146 Intron always much more larger than exon, despite exon is always more in number, not size.

DNA slippage : o Occurs in area where there are tandemly repeated sequences (e.g. STRshort tendom repeat – CACACACA) The complimentary code will be GTGTGTGT o There are thousands of STR in human genome o STR have a high rate mutation Hence knows as hot spots of mutation o Increase in repeat frequently are more common than decrease DNA is a very dynamic molecule, strands open and closes due to base pairing all the time, there will be an expansion in the DNA area, leading to elongation of the hot spot, which is known as DNA slippage, an alteration in the structure will take place, resulting in mutation. Somatic Mosaicism : Not all cells work 100% o Cells have genetic variation because :  Error in cell division  Error in replication  Random segregation of mitochondria - 2 places where u find DNA nucleous & mitochondria (mitochondrial DNA) - p.s. your mitochondria comes from your mother ½ cell only and the other half is in the tail of the sperm which does not fuse with the ova - its mutation due to( high metabolic rate in energy production therefore it requires different mitochondrial DNA so every cell has more than 100 mitochondria and when the cel is divided and it will form 2cells with different population of mitochondria and therefore different population of mitochondrial DNA) - when one mitochondria is not functioning and the other 99 is functioning you cannot see the disease cause it is masked :: ‫:: دعٌة نهتفكز فً حكًت ين اهلل ًجيهيا خهقو‬ ‫ فً انعانى نكنج نقٍج سبعت أنٌاع فقط ٌعنً حنا‬mitochondria DNA ‫نٌ جًعج كم‬ !! ‫جٍنا ين سبعت أيياث‬ ‫ًىذي يغانطت عهًٍت نًا ًرد فً انكتب انسًاًٌت فكهنا ٌعهى أننا ين آدو عهٍو انسالو‬ .‫ًحٌاء‬  Change in gene expression control Somatic variation is normal but is frequently masked. …………………………………….. #END#