Beruflich Dokumente
Kultur Dokumente
RNA Splicing:
http://highered.mheducation.com/sites/0072995246/student_view0/
chapter14/rna_splicing.html
Splicing
• The average human gene
occupies 27 kb of genomic DNA,
yet only 1.3 kb (~ 5 %) is used to
encode amino acids
• The correlation between exons
and domains found in some
genes suggests that the genes
were originally assembled from
smaller pieces
• The model of protein evolution
through the combination of
different exons is called the exon
shuffle model
Translation
• The mRNA is translated in the 5' -to-3' direction. The
polypeptide is synthesized from the amino end toward the
carboxyl end
Translation
The translation system consists
of five major components:
1. Messenger RNA: coding
sequence of bases
2. Ribosomes on which
protein synthesis takes
place
3. Transfer RNA translates
codons into amino acid
4. Aminoacyl-tRNA
synthetases: catalyzes
attachment of an amino
acid to its corresponding
tRNA molecule
5. Initiation, elongation, How Translation Works: http://highered.mcgraw-
hill.com/sites/0072995246/student_view0/chapter18/ho
and termination factors w_translation_works.html
Translation: Initiation
In eukaryotes, initiation takes
place by scanning the mRNA for
an initiation codon (AUG)
1. eIF4F binds to the 5' cap on
the mRNA and recruits eIF4A
and eIF4B
2. This creates a binding site for
a charged tRNAMet (an initiator
tRNA), bound with eIF2, and a
small 40S ribosomal subunit
together with eIF3 and eIF5
3. These components all come
together at the 5' cap and form
the 48S initiation complex
Translation: Initiation
4. Initiation complex moves along
mRNA in 3' direction, scanning for
the first AUG
5. eIF5 causes release of initiation
factors and recruitment of a large
60S ribosomal subunit
6. This subunit includes binding sites
for tRNA molecules: the E (exit)
site, the P (peptidyl) site, and the
A (aminoacyl) site.
7. tRNAMet is located in the P site
and the A site is the next in line to
be occupied.
8. tRNA binding is accomplished by
hydrogen bonding between the
AUG codon in the mRNA and the
three-base anticodon in the tRNA.
Translation: Elongation
1. The first step: the 40S subunit moves one codon farther
along the mRNA, and the charged tRNA corresponding to the
new codon is brought into the A site on the 60S subunit
2. A peptidyl transferase catalyzes a reaction in which the
bond connecting the methionine to the tRNAMet is transferred
to the amino group of the next amino acid, forming the first
peptide bond
Translation: Elongation
3. The next step: the 60S subunit swings forward to catch
up with the 40S, and simultaneously the tRNAs in the P
and A sites of the large subunit are shifted to the E and P
sites, respectively
Translation: Elongation
4. One cycle of elongation is now completed, and the entire
procedure is repeated for the next codon.
• Eukaryotes synthesize a polypeptide chain at the rate of
about 15 amino acids per second
• Elongation in prokaryotes is a little faster (about 20 amino
acids per second), but the essential processes are very
similar
Translation: Termination
• When a stop codon is
encountered, the tRNA holding
the polypeptide remains in the
P site, and a release factor
(RF) binds with the ribosome.
• GTP hydrolysis provides the
energy to cleave the
polypeptide from the tRNA to
which it is attached
• The 40S and 60S subunits are
recycled to initiate translation
of another mRNA
Translation
• Most polypeptides fold
correctly upon exiting
ribosome: they pass through
a tunnel in large ribosomal
subunit that is 35 amino acids
• Emerging from tunnel, protein
enters into a protein cradle
where it folds
• The proper folding of more
complex polypeptides is
aided by proteins called
chaperones and
chaperonins
Translation: Prokaryotes
1. In prokaryotes, mRNA molecules have no cap and there is no
scanning mechanism
2. In E. coli, IF-1 and IF-3 initiation factors interact with the 30S
subunit and IF-2 binds with a special tRNA charged with
formylmethionine tRNAfMet
3. These components bind with an mRNA at the ribosome-
binding site, RBS or the Shine–Dalgarno sequence.
Together, they recruit a 50S sub-unit
4. mRNA molecules contain information for the amino acid
sequences of several different proteins; such a molecule is
called a polycistronic mRNA
5. Cistron: DNA sequence that encodes a single polypeptide
chain
6. In a polycistronic mRNA, each protein coding region is
preceded by its own ribosome-binding site and AUG initiation
codon Initiation: http://highered.mcgraw-
hill.com/sites/0072995246/student_view0/chapter17/translation_initiation.html
Translation: Prokaryotes
• After the synthesis of one polypeptide is finished, the next
along the way is translated
• The genes contained in a polycistronic mRNA often encode
the different proteins of a metabolic pathway.
Elongation: http://highered.mcgraw-
hill.com/sites/0072995246/student_vi
ew0/chapter18/translation_elongatio
n.html
Termination: http://highered.mcgraw-
hill.com/sites/0072995246/student_vi
ew0/chapter18/translation_terminatio
n.html
Genetic Code
The genetic code is the list of all
codons and the amino acids that
they encode
1. Translation starts from a
fixed point and a single
reading frame maintained
throughout the process of
translation
2. Genetic code is universal: the same triplet codons specify
the same amino acids in all species
3. Each codon consists of three nucleotides.
4. Code is non-overlapping
5. Code is degenerate: each amino acid is specified by more
than one codon
Mutation
• Mutations occur when changes in codons alter amino
acids in proteins
• Mutations that delete or add a base pair shift the reading
frame and are called frameshift mutations.
How Is Gene Expression Regulated?
1. Cells can control the
frequency at which an
individual gene is transcribed
2. The same gene may be used
to produce different mRNAs
and protein products
3. Cells may control the stability
and translation of messenger
RNAs
4. Proteins may require
modification before they can
carry out their functions
5. Cells can control the rate at
which proteins are degraded
Epigenetics
1. Certain enzymes in a cell
add methyl groups (CH3)
to cytosine bases during a
process called methylation
http://www.bozemanscience.com/epigenetics