Molecular Biology and Genetics Notes 09/19/2016 Chapter 7

Anatomy and Function of a gene: Dissection through Mutation

1. Mutations: Primary tools of genetic analysis

A. Mutations are changes in DNA base sequences
I. Forward Mutation: changes wild type allele to a different allele
a. A+ - a or B+ - b
II. Substitution – replacement of a base by another base
a. Transition – purine replaced by another purine
b. Transversion – purine replaced by pyrimidine
III. Deletion – block of one or more bp lost
IV. Insertion – the addition of one or more nucleotide pairs
B. Spontaneous Mutations occur at a very low rate
I. Different genes, different mutation rates
a. Mutation rates are <10^-9 to >10^-3 per gene gamete
2. Molecular mechanisms of mutation
A. Natural Processes cause spontaneous mutations through DNA damage
I. Depurination – the hydrolysis of a purine base from the deoxyribose phosphate
backbone
a. Happens 1000/hr in every cell
II. Deamination – the removal of an amino group which can turn cytosine to uracil.
III. X-rays break the sugar – phosphate backbone of DNA
IV. Ultra-violet light (UV) causes adjacent thymine to form abnormal covalent bonds
(thymine dimers)
V. Oxidative damage
a. 8-oxodG mispairs with A
b. Normal C-G> mutant T-A after replication
V. Proofreading decreases mistakes made during replication
a. Incorporation of incorrect bases by DNA polymerase is exceedingly rare (<10^9)
in bacteria and humans)
i. Proofreading function of DNA polymerase – 3” to 5” exonuclease
recognizes and excises mismatches
VI. Base tautomerization results in replication mistakes
a. Rare tautomeric forms of bases have altered base pairing ability
b. Tautomerization causes single base pair mutations
VII. Expansion and contraction of triplet repeats occurs by slipped mispairing
a. Slipped mispairing – slippage of one DNA strand relative to the other during
DNA replication through repeat regions.
i. Trinucleotide repeat expansion – DNA polymerase pauses and newly
synthesized strand slips
ii. Tri nucleotide repeat contraction – DNA polymerase pauses and
template strand slips
B. The race between DNA repair and DNA replication
I. Base changes are often corrected by DNA repair.
 II. After replication, both DNA strands contain the base change and a heritable
mutation results
C. Mutagens Induce Mutations
I. Mutagen – anything that has a chemical damage on DNA
II. How mutagens alter DNA – Chemical actions of mutagen
a. Replace a base: base analogs – chemical structure almost identical to normal
base
b. Alter base structure and properties and properties: hydroxylating agents add an
-OH group.
c. Alter base structure and properties: Alkylating agents add ethyl or methyl
groups
d. Insert between bases: intercalating agents
III. Base excision repair removes damage bases
a. DNA glycosylases remove altered nitrogenous base
b. Nearby nucleotides are removes
c. New DNA is synthesized to fill gap
d. Particularly important for removing uracil (created by cytosine deamination)
from DNA
3. What mutations tell us about gene structure
4. What mutations tell us about gene function
5. A comprehensive example: Mutations that affect vision