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synthesis
Learning objectives
By the end of the lecture:
Students are expected to
understand the principles of gene
expression
students are expected to
understand and able to describe
the molecular or biochemical
processes underlying protein
synthesis
Core topics
Overview of Gene expression
Transcription of DNA (RNA
synthesis)
Protein synthesis (translation)
Overview of gene
expression
www.austincc.edu/.../1308ProteinSynthesis.pp
Cont.
Polypeptide/proteins are formed by
sequential addition of amino acids in
the specific order determined by info
carried in the nucleotide sequence of
the mRNA
Proteins are often matured or
processed by a variety of
modifications
Levels of translation is regulated
Transcription of DNA
(RNA synthesis)
mRNA
Carrier of information present in DNA
In eukaryotes (including human)
usually are synthesized as larger
precursor molecules that are
processed prior to export from the
nucleus
almost always monocistronic
(encoding a single polypeptide)
rRNA
Together with ribosomal proteins
constitute ribosome which in turn
serve as workbenches for
polypeptide/protein biosynthesis
Ribosome have two dissimilar
subunits (small & large subunits),
each contains RNA & many proteins
tRNA
A bilingual translator molecule
All tRNA molecules have several
common structural characteristics
(3-terminal CCA sequence to bind
amino acid, a highly conserved
cloverleaf secondary structure & Lshape three dimensional structure)
Great specificity in interaction with
mRNA & the aminoacyl-tRNA
synthetase
Involved:
RNA polymerase
NTP (ATP, GTP, CTP, UTP)
Termination signal
In most mammalian cells, only 1% of
the DNA sequence is copied into a
functional RNA (mRNA). Only one part
of the DNA is transcribed to produce
nuclear RNA, and only a minor portion
of the nuclear RNA survives the RNA
processing steps.
Promoter
Bind RNA polymerase protect DNA
from digestion
Two common motifs on 5 : -10
sequence 5-TATAAT-3 and -35
sequence (6 bp long) 5-TTGACA-3
At coding strand = sense (+) strand
& template strand = antisense (-)
strand
http://www.tokresource.org/tok_classes/biobiobio/biomenu/transcription_
RNA polymerase
Searches DNA for initiation site
There are many more molecules of RNA
polymerase per cell than DNA
polymerase.
RNA polymerase proceeds at a rate
much slower than DNA polymerase
(approximately 50-100 bases/sec for
RNA versus near 1000 bases/sec for
DNA
the fidelity of RNA polymerization is
DNA template
Transcription bubble for elongation
containing RNA pol, DNA, nascent
RNA
Form RNA-DNA hybrid helix (about 12
bp long/one turn of A-DNA)
Direct RNA synthesis
Transcribed by RNA pol (lack
nuclease activity) with lower fidelity
than that of replication (error rate 1
in 104 or 105)
Transcription termination
Processing of mRNA:
Capping at 5-end
Polyadenylation (tailing) at 3-end
Splicing removal of introns transcript
leaving exons transcript
http://vizonline.visitationacademy.org/upperSchool/essig/A
_gen.htm
Protein synthesis
(Translation of mRNA)
In eukaryotic cells,
translation is carried
out within cytoplasm
http://kentsimmons.uwinnipeg.
ca/cm1504/proteinsynth.htm
Translation of RNA
The ribosome binds to the mRNA at
the start codon (AUG) that is
recognized only by the initiator tRNA.
The ribosome proceeds to the
elongation phase of protein
synthesis. During this stage,
complexes, composed of an amino
acid linked to tRNA, sequentially bind
to the appropriate codon in mRNA by
forming complementary base pairs
with the tRNA anticodon.
tRNA molecule
http://kentsimmons.uwinnipeg.ca/cm1504/proteinsynth
.htm
Activation of amino
acid
Codon
Three-letter code words ( a triplet
code)
Unambiguous
Non-overlapping
Without punctuation
Universal
Can be found either in DNA (sense
strand) and mRNA
The collection of codons (64) makes up the genetic code
U
UUU
UUC
UUA
UUG
Phe
Phe
Leu
Leu
C
UCU
UCC
UCA
UCG
CUU
CUC
CUA
CUG
Leu
Leu
Leu
Leu
CCU
CCC
CCA
CCG
Pro
Pro
Pro
Pro
CAU
CAC
CAA
CAG
Tyr
Tyr
Stop
Stop
His
His
Gln
Gln
Ile
Ile
Ile
Met*
Val
Val
Val
Val
ACU
ACC
ACA
ACG
GCU
GCC
GCA
GCG
Thr
Thr
Thr
Thr
Ala
Ala
Ala
Ala
AAU
AAC
AAA
AAG
GAU
GAC
GAA
GAG
Asn
Asn
Lys
Lys
Asp
Asp
Glu
Glu
AUU
A AUC
AUA
AUG
GUU
G GUC
GUA
GUG
A
Ser
Ser
Ser
Ser
UAU
UAC
UAA
UAG
G
UGU
UGC
UGA
UGG
3'
Cys
Cys
Stop
Trp
U
C
A
G
CGU
CGC
CGA
CGG
Arg
Arg
Arg
Arg
U
C
A
G
AGU
AGC
AGA
AGG
GGU
GGC
GGA
GGG
Ser
Ser
Arg
Arg
Gly
Gly
Gly
Gly
U
C
A
G
U
C
A
G
Three
nonsense
codons (UAA,
UAG, UGA) do
not code for
specific amino
acid and are
utilized as
termination
signal
Cys:
Cysteine
Phe:
Phenylalanine Gly: Glycine
Lys: Lysine
Met:
Leu: Leucine
Methionine
Asn:
Asparagine
Pro: Proline
Gln:
Glutamine
Arg: Arginine
Ser: Serine
Thr:
Threonine
Val: Valine
Trp:
Tryptophane
Tyr: Tyrosisne
Polypeptide Elongation
Elongation
Initiation
Elongation
Termination
Mutations
Mutations
Result when changes occur in the
nucleotide sequence may not
occur in the template strand but
appear after replication
Some mutations occur by base
substitution single base changes
(point mutations):
Transitions (pryrimidine to other
pyrimidine, purine to other purine)
Transversion (pyrimidine to purine
or purine to pyrimidine)
3+
Inhibition of protein
synthesis
http://classes.midlandstech.com/carterp/Courses/bio225/chap
20/ss2.htm
http://classes.midlandstech.com/carterp/Courses/bio225/chap
20/ss2.htm