4/5/2017

What you need to know:

The key terms: gene expression, transcription, and
translation.
The major events of transcription.
How eukaryotic cells modify RNA after transcription. Concept 17.1:
The steps to translation. Genes specify proteins via
How point mutations can change the amino acid transcription and translation
sequence of a protein.

Gene Expression: process by which DNA

directs the synthesis of proteins (or RNAs) Flow of genetic information
Old idea: one gene-one enzyme hypothesis
Proposed by Beadle & Tatum mutant mold
experiments
Central Dogma: DNA RNA protein
Function of a gene = dictate production of specific
Transcription: DNA RNA
enzyme
Translation: RNA protein
Ribosome = site of translation
Newer idea: one gene-one polypeptide hypothesis

Most accurate: one gene-one RNA molecule (which can be

translated into a polypeptide)

Flow of Genetic one gene = one RNA molecule

Information in
Prokaryotes vs. DNA RNA
Eukaryotes Nucleic acid composed of Nucleic acid composed of
nucleotides nucleotides
Double-stranded Single-stranded
Deoxyribose=sugar Ribose=sugar
Thymine Uracil
Template for individual Many different roles!

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The Genetic Code

RNA plays many roles in the cell
1. pre-mRNA=precursor to mRNA, newly transcribed and not For each gene, one
edited DNA strand is the
2. mRNA= the edited version; carries the code from DNA that template strand
specifies amino acids
3. tRNA= carries a specific amino acid to ribosome based on its
anticodon to mRNA codon
mRNA (5 3)
complementary to
4. rRNA= makes up 60% of the ribosome; site of protein synthesis template
5. snRNA=small nuclear RNA; part of a spliceosome. Has
structural and catalytic roles
6. srpRNA=a signal recognition particle that binds to signal mRNA triplets
peptides (codons) code for
7. RNAi= interference RNA; a regulatory molecule amino acids in
8. ribozyme= RNA molecule that functions as an enzyme polypeptide chain

The Genetic Code

64 different codon
combinations
Redundancy: 1+
codons code for each Concept 17.2:
of 20 AAs Transcription is the DNA-directed
Reading frame: groups synthesis of RNA
of 3 must be read in
correct groupings
This code is universal:
all life forms use the
same code.

Transcription 1. Initiation
Transcription unit: stretch of DNA that codes for a
polypeptide or RNA (eg. tRNA, rRNA) Bacteria: RNA
polymerase binds
directly to promoter
RNA polymerase:
in DNA
Separates DNA strands and transcribes mRNA
mRNA elongates in 5 3 direction
Uracil (U) replaces thymine (T) when pairing to
adenine (A)
Attaches to promoter (start of gene) and stops at
terminator (end of gene)

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1. Initiation 2. Elongation
Eukaryotes:
TATA box = DNA
sequence (TATAAAA)
upstream from
promoter
Transcription RNA polymerase
factors must adds RNA nucleotides
recognize TATA box to the 3 end of the
before RNA growing chain (A-U, G-
polymerase can C)
bind to DNA
promoter

2. Elongation 3. Termination
RNA polymerase
transcribes a terminator
sequence in DNA, then
As RNA polymerase mRNA and polymerase
moves, it untwists DNA, detach.
then rewinds it after
mRNA is made It is now called pre-mRNA
for eukaryotes.

Prokaryotes = mRNA ready

for use

Flow of Genetic
Information in
Prokaryotes vs.
Eukaryotes

DNALC Video: Transcription

http://www.youtube.com/watch?v=SMtWvDbfHLo

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Additions to pre-mRNA:
5 cap (modified guanine) and 3 poly-A tail (50-
520 As) are added

Concept 17.3:
Eukaryotic cells modify RNA after
transcription

Help export from nucleus, protect from enzyme

degradation, attach to ribosomes

RNA Splicing RNA Splicing

small nuclear ribonucleoproteins
= snRNPs
snRNP = snRNA + protein
Pronounced snurps
Recognize splice sites
snRNPs join with other proteins
to form a spliceosome

Pre-mRNA has introns (noncoding sequences) and Spliceosomes catalyze the

exons (codes for amino acids) process of removing introns and
Splicing = introns cut out, exons joined together joining exons
Ribozyme = RNA acts as enzyme

Why have introns?

Some regulate gene activity

Alternative RNA Splicing:

produce different Concept 17.4:
combinations of exons Translation is the RNA-directed
One gene can make more synthesis of a polypeptide
than one polypeptide!
20,000 genes 100,000
polypeptides

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Components of Translation tRNA

1. mRNA = message Transcribed in nucleus
2. tRNA = interpreter Specific to each amino acid
3. Ribosome = site of translation Transfer AA to ribosomes
Anticodon: pairs with
complementary mRNA codon
Base-pairing rules between 3rd
base of codon & anticodon are
not as strict. This is called
wobble.

tRNA Ribosomes
Aminoacyl-tRNA-synthetase: Ribosome = rRNA + proteins
enzyme that binds tRNA to made in nucleolus
specific amino acid 2 subunits

Translation:
Ribosomes 1. Initiation

Active sites:
A site: holds AA to be added
P site: holds growing polypeptide
chain
E site: exit site for tRNA

Small subunit binds to start codon (AUG) on mRNA

tRNA carrying Met attaches to P site
Large subunit attaches

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2. Elongation 2. Elongation

Codon recognition:
tRNA anticodon
matches codon in A
site

2. Elongation 2. Elongation

Translocation: tRNA
in A site moves to
Peptide bond
P site; tRNA in P
formation: AA in A
site moves to E site
site forms bond with
(then exits)
peptide in P site

2. Elongation
3.Termination
Repeat over Stop codon reached and translation stops
and over Release factor binds to stop codon; polypeptide is
released
Ribosomal subunits dissociate

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Polyribosomes Protein Folding

A single mRNA can be During synthesis, polypeptide chain coils and folds
translated by several spontaneously
ribosomes at the same Chaperonin: protein that helps polypeptide fold
time correctly

Types of Ribosomes Cellular Zip Codes

Free ribosomes: synthesize proteins that stay in cytosol Signal peptide: 20 AA at leading end of polypeptide
and function there determines destination
Bound ribosomes (to ER): make proteins of Signal-recognition particle (SRP): brings ribosome to ER
endomembrane system (nuclear envelope, ER, Golgi,
lysosomes, vacuoles, plasma membrane) & proteins for
secretion
Uses signal peptide to target location

The Central Dogma

Mutations happen here

Concept 17.5:
Point mutations can affect protein
structure and function

Effects play out here

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Mutations = changes in the genetic

material of a cell Substitution = Missense
Large scale mutations: chromosomal; always cause
disorders or death
nondisjunction, translocation, inversions,
duplications, large deletions
Point mutations: alter 1 base pair of a gene
1. Base-pair substitutions replace 1 with another
Missense: different amino acid
Nonsense: stop codon, not amino acid
2. Frameshift mRNA read incorrectly; nonfunctional
proteins
Caused by insertions or deletions

Substitution = Nonsense Substitution = Silent (no effect)

Deletion = Extensive missense, premature

Insertion = Frameshift Mutation termination

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Sickle Cell Disease Sickle-Cell Disease = Point Mutation

Symptoms
Anemia Caused by a
Pain genetic defect
Frequent infections
Delayed growth Carried by 5% of
Stroke humans
Pulmonary hypertension
Organ damage Carried by up to
Blindness 25% in some
Jaundice regions of Africa
gallstones
Life expectancy
42 in males 48 in females

Sickle cell hemoglobin forms long,

inflexible chains

Comparison:
Prokaryotes vs. Eukaryotes

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Prokaryote vs. Eukaryote Prokaryotes vs. Eukaryotes

Prokaryotes
Transcription and translation
both in cytoplasm
DNA/RNA in cytoplasm
RNA poly binds directly to
promoter
Transcription makes mRNA
(not processed)
No introns
Eukaryotes
Transcription in nucleus;
translation in cytoplasm
DNA in nucleus, RNA
travels in/out nucleus
RNA poly binds to TATA
box & transcription factors
Transcription makes pre-
mRNA RNA processing
final mRNA
Exons, introns (cut out)

A Summary
of Protein
Synthesis
(p. 348)

Most current
definition for a
gene: A region of
DNA whose final
product is either a
polypeptide or an
RNA molecule

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