Mutation

A sudden & heritable change in genetic material. New source of variation (departure from Wild Type) - mutant Has many forms & various mechanisms mutational changes in genotype of an organism include changes in CHROMOSOMAL NUMBERS & STRUCTURE (gross mutation) and changes in STRUCTURES OF INDIVIDUAL GENES (fine-scale mutation) - Today? Mutation refers to changes that occur within genes Factors of mutation? Provide raw material for evolution. (Mutation is source of all genetic variation). Gives new combination (Recombination mechanism rearrange genetic variability) For natural or artificial selection (esp. plant & animal breeder) For organisms to adapt to environmental changes. Basic Features of the Process? Type of cell: Somatic / Germinal Ways? Spontaneous / Induced a random, nonadaptive process a reversible process Somatic Mutation Occur in somatic cell Mutant phenotype occur only in the descendants of the cell Not transmitted through the gametes to the progeny Spontaneous mutation occur without a known cause - maybe resulting from a low level of inherent metabolic error / unknown agents present in the environment Definition Germinal Mutation Occur in germ-line cells Occur in any stage in reproductive cycle; either dominant / recessive. Immediate effect or obscured in diploids.

Induced mutation resulting from exposure of organisms to physical/chemical agents that causes changes in DNA (or RNA in some viruses)

Spontaneous mutation tautomeric shift Molecular basis of mutation

Induced mutation chemicals radiation transposable genetic element

Spontaneous Mutation
Mechanism How? Non-disjunction of homologous chromosomes Random during meiosis 1 / chromosome meiosis 2 breaks proposed by Watson & Crick. 4 organic bases in nucleotides (A,G,C,T) are not 100% stable & may pair up with noncomplementary bases. eg? normally, A will pair with T, but occasionally pair up with G the

3 I. II. III.

Tautomeric shifts; alter pairing of bases. mechanism of tautomeric shifts:

Transitions Transversion Frameshift mutations Occur at the rate of 10-6 or 10-8 replications. Causes?

Induced Mutation
Agents that cause mutation? Mutagents. Caused by exposure of DNA to mutagens Examples of mutagens:

Chemicals

Radiation

Transposable genetic element

sources of exogenous DNA damage: Natural: aflatoxin (in food) Man-made: -Nitrogen Mustard (WW1 nerve gas). -Benzopyrene (smoke from coal, cigs, automobiles) Chemical Mutagens Nitrous oxide & HNO2 Deaminates C & changes it to U 5-bromouracil (base analogs) Incoporates itself into nucleic acids in place of one of normal bases e.g: 5-bromouracil pairs with guanine in place of adenine Mustard gas Replace G in DNA with other bases Caffeine Weak mutagen - base analog of A & sometimes can take As place in DNA UV radiation (from Sun) Ionizing radiation: Natural: Rd gas, cosmic ray Manmade:xray, nuclear test

Cyclamate cause chromosomal aberrations Acridine orange addition / removal of bases in DNA

Nitrous oxide & HNO2 Deaminates C & changes it to U

5-bromouracil (base analogs) Incoporates itself into nucleic acids in place of one of normal bases e.g: 5-bromouracil pairs with guanine in place of adenine Ethyl methanesulfonate chemical mutagens that react with bases and add methyl or ethyl groups. Depending on the affected atom, the alkylated base may then degrade to yield a baseless site, which is mutagenic and recombinogenic, or mispair to result in mutations upon DNA replication. Ultraviolet Radiation

Formaldehyde Mutagenic - cause gene mutations & chromosomal aberrations when came in contact with cells

Physical mutagens Ionizing Radiation: X-rays & gamma rays Cause simple base substititions Produce large deletion of gen material due to breaks produced as a result of collisions between high energy particles & DNA molecule.

Cause abnormal bonding of T base Inactivates a DNA strand as dimer cannot form H bonds with A nucleotides of the complementary strand. Thus, DNA replication & mRNA transcription cannot occur.

Nitrous acid mutations

Uses of Mutation?
Spontaneous mutations sometimes produce useful mutations in plants cultivated by man, but occur at a very low rate & takes a long time to produce beneficial results. Induced mutation using chems & radiation can produce mutation that increase yeilds & resistance in crops including cereals, oil-producing plants, root & tuber plants.

Use of induced mutation in plant improvement?

Can use chems like EMS, DMSO/ colchine/ radiation. using radiation, seeds are irradiated & allowed to grow to seedlings. Then, they are selected for favorable traits. Eg of plants produced by radiation: Barley that can grow 5000m above sea level. Rice can grow in high salinity water.

Gene Mutation (Fine-Scale Mutations)

Involve less than 1000 base pairs Involve point mutation (just a single base pair is changed)

Include SUBSTITUTIONS, DELETIONS, INSERTIONS, DUPLICATIONS Normal: 5-------GAATTC-------3 3-------CTTAAG-------5 Insertion 5---GAACTTC---3 3---CTTGAAG---5 Duplication 5---GAATATTC---3 3---CTTATAAG---5

Substitutions Occur in protein-coding sequences

Deletion/insertion Occur in protein-coding sequences in multiples other than 3 (cause frameshift mutations) FrAmEsHiFt eX: Insertion NH2 - Met - Thr - Leu - Lys - COOH 5- ATG-ACC- TTG - AAA - TAA - 3 Deletion NH2 - Met - Pro - COOH 5-ATG-CCT-TGA-AAT-AA-3

Silent - changes a codon, but not the encoded amino acid residue. Possible as the code is degenerated. TGT(Cys) ------> TGC(Cys) GCA(Ala) ------> GCN (Ala) (N=any base) Missense - changes the encoded residue TGT(Cys) -------> TGG(Trp) Nonsense - an amino acid-encoding codon becomes a stop codon TGT(Cys) --------> TGA(Stop codon)

Mutation - Case study Mutation: produces blocks in metabolic pathway, because alterations in the base-pair sequences of the gene, causes changes in amino acid sequence of polypeptides, causes polypeptides to become a nonfunctional product. Examples: Sickle-cell hemoglobin Phenylketonuria Tay-Sachs disease

Chromosomal Mutation (Gross Mutation)

Observable cytogentically (at level of the chromosome) Include DELETIONS, DUPLICATIONS, INVERSIONS, TRANSLOCATIONS May involve: Changes in amount & arrangement of chromosomes (chromosome aberration) Changes in chromosome number. Chromosomal Aberrations:
Translocation Involves a region of a chromosome breaking off & rejoining one end of the same chromosome or another non-homologous chromosome. Some chromosomes have centromeres located near one end so that one arm is long acrocentric chromosomes. 5 of acrocentric chromosomes: 13, 14, 15, 21, 22. Sometimes, the long arms of the chromosomes may come together & fuse, and transfer of chromosomes may occur from one chromosome to another; the transfer of chromosome segments are called Robertsonian translocation. Most frequent are between chromosomes 13 and 14, 13 and 21, 21 and 22. Advantages? Some translocation may result in rapid evolutionary changes. Disadvantages? -May activate genes that can lead to cancer & reduced fertility. -May lead to loss of important chromosome segments. ABCDEFGHIJK --------> Normal ABCDEFGHIJKXYZ----> Additional genes from other chromosome.

Deletion Involves loss of segment from the middle/ the end of a chromosome. This segment does not get fused with other chromosome but entirely lost. Advantages? Loss of unfavorable genes. Disadvantages? -Loss of some gene maybe lethal to the individual. -Disrupt chromosomal disjunction during meiosis. ABCDEFGHIJK --------> Normal ABCDEIJK--------------> FGH segment is lost Inversion Occurs when a region of chromosome breaks off & rotates through 180 before being reattached back into the chromosome in reverse order. Advantages? May increase gen diversity by changing gene loci Disadvantages? Reduce fertilty; loss of control of gene expression ABCDEFGHIJK --------> Normal ABFEDCGHIJK --------> CDEF is inverted, becomes FEDC Duplication Sometimes, a particular segment of chromosome may become duplicated so that some genes on a chromosome may be repeated. Advantages? -Provide back up genes to cover needs if mutation inactivates a gene -Increase gene dosage to boost production of some products Disadvantages? -May interfere with chromosome separation -May disrupt gene func if duplication occurs within a gene ABCDEFGHIJK ---------> Normal ABCABCDEFGHIJK ----> Genes ABC repeated

Changes in chromosomal number

2 types of change in chromosomal number: Aneuploidy Loss/gain of 1 chromosome. Caused by nondisjunction during meiosis 1 / meiosis 2 nondisjunction can occur in both autosomes & sex chromosomes. Polyploidy Increase in the whole set of haploid chromosomes, eg: 3n, 4n.

Aneuploidy
If nondisjunction occurs during meiosis 1, the resulting gametes produced is (n+1) or (n-1). If nondisjunction occur during meiosis 2, the resulting gametes produced is (n+1) or (n) or (n-1). Condition of (n+1) chromosomes: Down syndrome (trisomy 21) Patau syndrome (trisomy 13) Klinefelter syndrome (XXY) Condition of (n-1) chromosomes: Monosomy 21 Turner syndrome (XO)

1. Nondisjunction during meiosis

Spindle fibre fails to form during meiosis

Members of one pair of homologous chromosomes fail to separate Thus, 2 gametes receive extra copy of affected chromosome 2 gametes lack that chromosome Patau syndrome (Trisomy 13) Low birth weight Structural eye defects Cleft palate & hare lip Extra digits & overlapping of fingers over thumb Death by 1-3 months

Edwards syndrome (Trisomy 18) Ear deformities Heart defects Spasticity Death by 1 year

Down syndrome (Trisomy 21) A fold of skin above the eye Short stature

Protruding tongue Cardiac deformities Varying degrees of mental retardation Develop slower have heart & stomach illness vary in level of intelligence Why? Nondisjunction in chromosome 21 Occur in egg mother cell One/more abnormal eggs formed (n=24) Fertilized by normal sperm (n=3) Form abnormal zygote (2n=47)

2.
Turner Syndrome Female

Nondisjunction of sex chromosomes

Short in stature Infertile as ovaries does not develop normally Webbed neck ovaries degenerate rudimentary sexual characteristics Why? Gametes with no sex chromosomes fuses with X gamete Thus, zygote has chromosome complement 2n=45 Klinefelters syndrome (XXY) Male Normally infertile Cannot produce sperm Testes degenerate Enlarge breasts Why? XX egg fertilized by normal Y sperm OR Normal X egg fertilized by XY sperm Thus, zygote has chromosome complement 2n=47 (44+XXY)

XXX syndrome Female Little/no visible differences tall stature learning disabilities limited fertility XYY Syndrome Normal male traits Often tall & thin Associated with antisocial & behavior problems

3.

Complete non-disjunction & polyploidy

Fertilization of abnormal gametes Formation of mutant plants that posess extra sets of chromosomes

Why? All spindle fibres in a gamete mother cell fail to form All homologous chromosome pairs fail to be separated Thus, Abnormal diploid gametes is produced Each contain 2 complete sets of chromosomes instead of one Autopolyploidy 2n diploidy 3n triploidy 4n tetraploidy 5n pentaploidy 6n polyploidy Caused by colchicine which disrupts the microtubules so that the chromatids do not separate during anaphase. In mitosis, if cytoplasm fails to cleave & divide during cytokinesis, a tetraploid cell (4n) with a large nucleus is produced. Tetraploid cell may be produced in the liver when chromosome replication occurs without cell division. Allopolyploidy 2(A+B) x 2(A+B) crossed

F1 hybrids produced from diff species are sterile as there are no homologous chromosomes & pairing cannot occur during meiosis. This is hybrid sterility. If multiples of original haploid no. of chromosomes, eg: 2(A+B), 4(A+B) & so on (A & B are the haploid no. of the parent species) occur, a new species is produced which is fertile when crossed to polyploids like itself (hybrid) but sterile when crossed to parent species.

Economic significance Polyploid plants are larger than diploid relatives. Increased seed & fruit size. Give greater yeilds. Comercially developed cropped plants: Strawberries Apples Wheat Tomatoes Effects of polyploidy Uneven sets of chromosome - polyploid plants are sterile Produce seedless fruit Increase in vigour Resistance to disease

A: strawberry plant is exposed to chemical

B: due to the effect of chemical, total nondisjunction at meiosis produces diploid gametes (2n) C: 2 diploid gametes fuse at fertilization

D: a new plant develops which has 4 sets of chromosomes (4n)

Spartina stricta (2n=60) AA

Spartina alternifora (2n=62) BB

Spartina Townsendii (sterile f1 hybrid) (2n=61) AB

Spartina Anglica (fertile allopolyploid)

(2n=122) AABB