Transcript AP Protein Synthesis

From DNA to Proteins
Steps from DNA to Proteins
Same two steps produce all proteins:
1) DNA is transcribed to form RNA
– Occurs in the nucleus
– RNA moves into cytoplasm
2) RNA is translated to form polypeptide
chains, which fold to form proteins
– occurs at the ribosome
Three Main Classes of RNAs
• Messenger RNA
– Carries protein-building instruction
• Ribosomal RNA
– Major component of ribosomes
• Transfer RNA
– Delivers amino acids to ribosomes
Differences Between DNA and RNA
1. Sugars – DNA = deoxyribose, RNA = ribose
2. DNA = double strand, RNA = single strand
3. DNA has Thymine as one base, RNA has
Uracil instead
A Nucleotide Subunit of RNA
uracil (base)
phosphate
group
sugar
(ribose)
Figure 14.2
Page 228
Base Pairing during
Transcription
DNA
G
C
A
T
RNA
G
C
A U
DNA
C
G
T
A
DNA
C
G
T A
base pairing in DNA replication
base pairing in transcription
Transcription & DNA
Replication
• Like DNA replication
– Nucleotides added in 5’ to 3’ direction
• Unlike DNA replication
– Only small stretch is template
– RNA polymerase catalyzes nucleotide
addition
– Product is a single strand of RNA
• A base sequence in the DNA that signals the start of a
gene (TATA)
• For transcription to occur, RNA polymerase must first
bind to a promoter (in eukaryotes this requires many
transcription factors)
• Once RNA polymerase is bound it will unwind the DNA
and nucleotides can be added
Gene Transcription
transcribed DNA
winds up again
DNA to be
transcribed unwinds
mRNA
transcript
RNA polymerase
Figure 14.4c
Page 229
Adding Nucleotides
5’
growing RNA transcript
3’
5’
3’
direction of transcription
Figure 14.4d
Page 229
Once the termination site is reached,
the RNA molecule will be released. In
eukaryotic cells, the RNA polymerase
will actually pass the termination point
before the RNA molecule is released.
Transcript Modification
unit of transcription in a DNA strand
3’
exon
intron
exon
transcription
intron
5’
exon
into pre-mRNA
poly-A
tail
3’
cap
5’
snipped
out
snipped
out
5’
3’
mature mRNA transcript
Figure 14.5
Page 229
RNA processing1. 5' cap with a modified guanine nucleotide is added.
2. At the 3' end 30-200 adenine nucleotides are added (poly-Atail).
-These modifications prevent the mRNA from being degraded and
signal the ribosome where to attach.
3. There are noncoding regions (introns) that are removed in
eukaryotic cells. The remaining regions (exons) are joined
together. A particle called a spliceosome removes the
introns. Spliceosomes are composed of smaller particles
called snRNP (made of proteins and snRNA).
-
The average immature RNA is 8000 nucleotides long but the
mature mRNA is 1200 nucleotides long.
Genetic Code
• Set of 64 base triplets
• Codon = 3 bases
• 61 specify amino acids.
There is "redundancy" in
the code or more than one
codon codes for the same
amino acid. One that
codes for methionine also
codes for “start”.
• 3 stop codons
Figure 14.7
Page 230
tRNA Structure
codon in mRNA
anticodon
amino-acid
attachment site
amino
acid
OH
Figure 14.8
Page 231
Ribosomes
tunnel
small ribosomal subunit large ribosomal subunit
intact ribosome
Figure 14.9b,c
Page 231
Three Stages of Translation
Initiation
Elongation
Termination
Initiation
• Initiator tRNA binds to small
ribosomal subunit
• Small subunit/tRNA
complex attaches to mRNA
and moves along it to an
AUG “start” codon
• Large ribosomal subunit
joins complex
Fig. 14.10a-c
Page 232
Elongation
• mRNA passes through ribosomal
subunits
• tRNAs deliver amino acids to the
ribosomal binding site in the order
specified by the mRNA
• Peptide bonds form between the amino
acids and the polypeptide chain grows
Elongation
Fig. 14.10e-g
Page 233
Termination
• Stop codon into place
• No tRNA with anticodon
• Release factors bind to
the ribosome
• mRNA and polypeptide
are released
mRNA
new
polypeptide
chain
Fig. 14.10j-k
Page 233
What Happens to the
New Polypeptides?
• Some just enter the cytoplasm
• Many enter the endoplasmic reticulum
lumen and move through the
cytomembrane system where they are
modified
Transcription
Overview
mRNA
Mature mRNA
transcripts
Translation
rRNA
ribosomal
subunits
tRNA
mature
tRNA
http://www.fed.cuhk.edu.hk/~johnson/teaching/
genetics/animations/transcription.htm
http://www.stolaf.edu/people/giannini/flashanim
at/molgenetics/translation.swf
http://www.youtube.com/watch?v=WsofH466lqk
Gene Mutations
Base-Pair Substitutions (point mutation)
Insertions (frameshift)
Deletions (frameshift)
Base-Pair Substitution
•Results in 1 wrong amino acid
•Protein may still function
a base substitution
within the triplet (red)
original base triplet
in a DNA strand
During replication, proofreading
enzymes make a substitution
possible outcomes:
or
original, unmutated
sequence
a gene mutation
Figure 14.11
Page 234
Frameshift Mutations
• Insertion
– Extra base added into gene region
• Deletion
– Base removed from gene region
• Both shift the reading frame
• Result in many wrong amino acids
Frameshift Mutation
mRNA
parental DNA
arginine
glycine
tyrosine
tryptophan asparagine amino acids
altered mRNA
arginine
glycine
leucine
leucine
glutamate
DNA with
base insertion
altered aminoacid sequence
Figure 14.12
Page 234
Transposons
• DNA segments that move
spontaneously about the genome
• When they insert into a gene region,
they usually inactivate that gene
Mutation Rates
• Each gene has a characteristic mutation
rate
• Average rate for eukaryotes is between
10-4 and 10-6 per gene per generation
• Only mutations that arise in germ cells
can be passed on to next generation
Mutagens – things that can lead
to mutations
• Ionizing radiation (X rays)
• Nonionizing radiation (UV)
• Natural and synthetic chemicals