Germline Somatic
Inherited Acquired
mutations mutations
Mutations and biotechnology
- Generating mutants - traditionally
selective breeding
inducing mutations (x-rays, chemicals)
. PHYSICAL
Radiation: extra energy changes stability
UV light
, , rays (think Chernobyl)
X-rays
Example: UV light causes Cs and Ts to dimerize
. CHEMICAL
Free radicals, tar, asbestos, charred steaks
. CHANCE
This chance is increased over multiple cycles of replication.
Cycles of replication are triggered by several causes
growth, repair, replenish (and some tissues are constantly
regenerating, like the G.I. epithelium)
Mutations: Causes and Consequences
Mutation change in the nucleotide sequence of DNA
Major cause of genetic diversity
Can also be detrimental
Types of Mutations
Point/Substitution mutations
Silent mutations
Missense mutations
Nonsense mutations
Frameshift mutations
Insertion or deletion mutations
viruses,
Transposable elements (jumping genes),
chromosomal translocations
Mutations: Types
Proofreading
Point mutation
Chromosomal mutations
Repair of DNA errors
Error rate of 1 for every 105 bases
replicated.
If uncorrected, ~60,000 mutations each
division
At least three mechanisms of repair
Proofreading
Mismatch repair
Excision repair
What Do Mutations Do?
DNA codes for proteins via transcription and
translation
A mutated mRNA transcript provides faulty
directions for protein formation
Different order of bases = different codons
As a result, the order or length of the amino
acid sequence may be changed
This affects protein SHAPE, and hence, its
function (remember?)
Mutations in non-translated RNA are also very
dangerous. Think, telomers, tRNA, rRNA
Mutations: Consequences
Chromatin Structure
double
helix
(2-nm
diameter)
Histones
DNA is packed, coiled, around Beads on
a string
histones, a kind of protein.
700
nm
Metaphase chromosome
Copyright 2003 Pearson Education, Inc. publishing as
Benjamin Cummings
Epigenetic Modifications
Methylation:
-CH3
Heterochromatin
Tight association
between histones
and DNA
Acetylation:
- CH2CO
Euchromatin
Loose association
between histones
and DNA
Transcriptional regulation
Controlling the amount of mRNA transcribed from
a particular gene
Conserved sequences in the promoter region affect the
affinity of RNA polymerase
TATA box at -35 (TATAAT - pribnow box in procaryotes)
CAAT box at -80
RNA polymerase cannot bind to promoter region without
presence of transcription factors
Enhancer sequences bind to regulatory regions called
activators
Upstream Activating Sequences are 50 bp or more
upstream or even downstream
Repressors feed back inhibition
Transcriptional Regulation
Transcription factors are themselves
proteins
So what regulates their expression?
1. Feedback loops
the presence or absence of the product the protein acts
upon may be a switch
Either all three genes are turned on, in the presence of lactose
Or all three genes are turned off in the absence of lactose
When lactose binds the repressor protein, its conformation changes and it can no
longer bind to the promoter thus inhibiting RNA polymerase to interact with DNA