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Transcript
Control of
Eukaryotic Genes
and
Epigenetics
AP Biology
2007-2008
The BIG Questions…
■
■
How are genes turned on & off
in eukaryotes?
How do cells with the same genes
differentiate to perform completely
different, specialized functions?
AP Biology
Evolution of gene regulation
■
Prokaryotes
single-celled
 evolved to grow & divide rapidly
 must respond quickly to changes in
external environment

■
■
exploit transient resources
Gene regulation

turn genes on & off rapidly
■

AP Biology
flexibility & reversibility
adjust levels of enzymes
for synthesis & digestion
Evolution of gene regulation
■
Eukaryotes
multicellular
 evolved to maintain constant internal
conditions while facing changing
external conditions

■

homeostasis
regulate body as a whole
■
growth & development
 long term processes
■
specialization
 turn on & off large number of genes
■
AP Biology
must coordinate the body as a whole rather
than serve the needs of individual cells
Points of control
■
The control of gene
expression can occur at any
step in the pathway from
gene to functional protein
packing/unpacking

1.
DNA
2.

3.
mRNA processing

4.
mRNA transport

5.
translation

6.
protein processing
7.
protein degradation

AP Biology

transcription
1. DNA packing
How do you fit all
that DNA into
nucleus?
■

DNA coiling & folding
■
■
■
■
■
■
■
double helix
nucleosomes
chromatin fiber
looped domains
chromosome
Link to Animation1
Link to Animation 2
from DNA
double helix to
AP Biology
condensed
Nucleosomes
■
8 histone
molecules
“Beads on a string”
1st level of DNA packing
 histone proteins

■
■
■
8 protein molecules
positively charged amino acids
bind tightly to negatively charged DNA
AP Biology
DNA
packing movie
DNA packing as gene control
■
Degree of packing of DNA regulates transcription

tightly wrapped around histones
■
■
no transcription
genes turned off
 heterochromatin
darker DNA (H) = tightly packed
 euchromatin
lighter DNA (E) = loosely packed
H
AP Biology
E
DNA methylation
■
Methylation of DNA blocks transcription factors

no transcription

 genes turned off
attachment of methyl groups (–CH3) to cytosine in
■
■

C = cytosine
nearly permanent inactivation of genes in females(XX)
■
■
■
AP Biology
ex. inactivated mammalian X chromosome = Barr body
why? X VERY LARGE..too much protein if both expressed.
Link to Animation (6:05)
Methylation a form of Genetic Imprinting (Memory) and silences
selected genes. “Pins” represent methyl group additions
AP Biology
10
Histone acetylation
■

■
■

■
■
■
■
Acetylation of histones unwinds DNA
loosely wrapped around histones
enables transcription
genes turned on
attachment of acetyl groups (–COCH3) to lysine amino acids within
the histone protein
conformational change in histone proteins
transcription factors have easier access to genes
Link to Animation
Link to Animation
AP Biology
2. Transcription initiation
■
Control regions on DNA

promoter
■
■
■

enhancer
■
■
■
AP Biology
nearby control sequence on DNA
binding of RNA polymerase & transcription
factors
“base” rate of transcription
distant control
sequences on DNA
binding of activator
proteins
“enhanced” rate (high level)
of transcription
Model for Enhancer action
■
Enhancer DNA sequences

■
Activator proteins

■
distant control sequences
bind to enhancer sequence &
stimulates transcription
Silencer proteins

bind to enhancer sequence &
block gene transcription
AP Biology
Turning
on Gene movie
Transcription complex
Activator Proteins
• regulatory proteins bind to DNA at
Enhancer Sites
distant enhancer sites
• increase the rate of transcription
regulatory sites on DNA
distant from gene
Enhancer
Activator
Activator
Activator
Coactivator
A
E
F
B
TFIID
RNA polymerase II
H
Core promoter
and initiation complex
Initiation Complex at Promoter Site binding site of RNA polymerase
AP Biology
3. Post-transcriptional control
■
Alternative RNA splicing

AP Biology
variable processing of exons creates a
family of proteins
4. Regulation of mRNA degradation
■
Life span of mRNA determines amount
of protein synthesis

mRNA can last from hours to weeks
AP Biology
RNA
processing movie
RNA interference
■
Small interfering RNAs (siRNA)

short segments of RNA (21-28 bases)
■
■
■
bind to mRNA
create sections of double-stranded mRNA
“death” tag for mRNA
 triggers degradation of mRNA

cause gene “silencing”
■
■
■
■
post-transcriptional control
turns off gene = no protein produced
Link to Animation 1
Link to Animation 2
siRNA
AP Biology
Action of siRNA
dicer
enzyme
mRNA for translation
siRNA
Text
double-stranded
miRNA + siRNA
breakdown
enzyme
(RISC)
Link to Animation
Link to Animation
mRNA degraded
AP Biology
functionally turns
gene off
RNA interference
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTi me™ a nd a
TIFF (Uncompre ssed ) decomp resso r
are need ed to se e th is p icture.
Andrew
Fire
AP
Biology
Stanford
QuickTi me™ a nd a
TIFF (Uncompre ssed ) decomp resso r
are need ed to se e th is p icture.
Craig Mello
U Mass
1990s | 2006
“for their discovery of
RNA interference —
gene silencing by
double-stranded RNA”
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
5. Control of translation
■
Block initiation of translation stage

regulatory proteins attach to 5' end of
mRNA
■
■
prevent attachment of ribosomal subunits &
initiator tRNA
block translation of mRNA to protein
AP Biology
Control
of translation movie
6-7. Protein processing & degradation
■
Protein processing

■
folding, cleaving, adding sugar groups,
targeting for transport
Protein degradation
ubiquitin tagging
 proteasome degradation

AP Biology
Protein
processing movie
1980s | 2004
Ubiquitin
■
“Death tag”
mark unwanted proteins with a label
 76 amino acid polypeptide, ubiquitin
 labeled proteins are broken down
rapidly in "waste disposers"

■
AP
proteasomes
Aaron Ciechanover
Biology Israel
Avram Hershko
Israel
Irwin Rose
UC Riverside
Proteasome
■
Protein-degrading “machine”


cell’s waste disposer
breaks down any proteins
into 7-9 amino acid fragments
■ cellular recycling of amino acids
■ Link to Animation
■ Link to Animation
AP Biology
play
Nobel animation
6
7
Gene Regulation
protein
processing &
degradation
1 & 2. transcription
- DNA packing
- transcription factors
(acetylation/methylation)
5
3 & 4. post-transcription
- mRNA processing
4
- splicing
mRNA
- 5’ cap & poly-A tail
processing
- breakdown by siRNA
initiation of
translation
5. translation
- block start of
translation
1 2
initiation of
transcription
AP Biology mRNA splicing
3
6 & 7. post-translation
- protein processing
- protein degradation
4
mRNA
protection
The Ghost in Your Genes Full Documentary on Epigenetics
Epigenetics Tutorial from NOVA
Epigenetics Tutorial from Cancer Institute
Turn your
Question Genes
on!
AP Biology
2007-2008
6
Gene Regulation
7
1 & 2. _________________
- ____________________
- ____________________
5
4
1 2
3 & 4. _________________
- ____________________
- ____________________
- ____________________
- ____________________
5. _________________
- ____________________
____________________
AP Biology
3
4
6 & 7. _________________
- ____________________
- ____________________