Central Dogma of Molecular

Biology
Steps from DNA to Proteins

Same two steps produce all proteins:

1) DNA is transcribed to form RNA
 Transcription
 Occurs
in the nucleus
 RNA moves into cytoplasm

2) RNA is translated to form polypeptide

chains, which fold to form proteins
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
Transcription
 DNA serves as a template for RNA synthesis
 Transfer of information from one nucleic acid to
another
 4 steps (initiation, elongation, termination and
posttranscriptional modification)

DNA G C A T RNA G C A U

DNA C G T DNA C G T A
A
base pairing in DNA replication base pairing in transcription
Transcription in
Prokaryotes
Initiation
- RNA polymerase bound to a promoter site and
locally unwound the DNA double helix
- Has a sigma (δ) factor which ensures initiation
at the proper sites within a DNA molecule
- one of the two single stranded DNA serves as
template for RNA synthesis
RNA Polymerase
 Catalyzes the
synthesis of
RNA using DNA
as a template
Three Classes of RNAs
 Messenger RNA
- Carries protein-building instruction
 Ribosomal RNA
- Major component of ribosomes
 Transfer RNA
- Delivers amino acids to ribosomes
Elongation
- RNA polymerase continues to
move along the DNA molecules and
to unwind helix
Termination
 RNA synthesis stops when RNA
polymerase copies a special sequence
called TERMINATION SIGNAL
- rho (ρ) factor
- Hairpin loop
Rho (ρ) factor
Hairpin loops
Transcription in Prokaryotes
Transcription in
Eukaryotes
Eukaryotes vs. Prokaryotes
 3 different RNA polymerases transcribe the nuclear DNA
of eukaryotes
 Eukaryotic promoters are more varied than prokaryotic
promoters.
 Binding of eukaryotic RNA polymerases to DNA requires
the participation of additional proteins called
TRANSCRIPTION FACTORS
 RNA cleavage is more important than the site when
transcription is terminated in determining the location of
the 3’ end of the RNA product
 Undergo further RNA processing
RNA Polymerase I
 Resides in the nucleolus
 Responsible for synthesizing an RNA
molecule that serves as a precursor for
three of the four types of rRNA (28S rRNA,
18S rRNA and 5.8 S rRNA)
RNA Polymerase II
 Found in nucleoplasm
 Synthesizes precursors of mRNA
 Synthesizes most of the snRNAs (small
nuclear RNAs)
RNA Polymerase III
 Found in nucleoplasm
 Synthesizes a variety of small RNAs
(tRNA precursors and the smallest type of
rRNA 5S rRNA)
Promoters
 RNA Polymerase II
-
Transcription factors (TF)
 Also called basal transcription factor
 Protein that is always required for an RNA
Polymerase molecule to bind to its promoter
and initiate RNA synthesis
 eg. TFIIA, TFIIB
Transcription Factors
TFIID – initial transcription
factor to bind to the
promoter (TATA box)
TFIIH – possesses both a
helicase activity and protein
kinase activity that catalyzes
the phosphorylation of RNA
polymerase II
Elongation
 RNA polymerase recruit proteins that unfold the higher-
order nucleosomal packaging of the chromatin fibers
that would otherwise block the enzyme from advancing
along the DNA template
Termination
 RNA Polymerase II
- most transcripts destined to become mRNA
are cleaved at a specific site before
transcription is actually terminated
RNA polymerase falls off
Post-transcriptional Modification
 All chemical
modifications
necessary to generate
a final RNA product
from the primary
transcript that serves
as its precursor
- capping
- addition of Poly(A)
- removal of introns
5’ Caps
 5’ cap is a guanosine
nucleotide that has
been methylated at
position 7 of the
purine ring
 Contributes mRNA
stability by protecting
the molecule from
degradation by
nucleases that attack
the RNA at the 5’ end
Poly(A) tail (Polyadenylation)
 Present at 3’ end of
eukaryotic mRNA
molecules
 Consists of adenine
bases
 Protects mRNA from
nuclease attack
 Stability (the longer the
tail, the longer the life
span of the mRNA in the
cytoplasm)
Introns
 Intervening sequences
 Sequences within the
primary transcript that do
not appear in the mature,
functional RNA
 Characterized by GU and
AG sequences
 Counterpart of exons
RNA Splicing
 Removal of introns and
rejoining exons
 RNA splicing must be
very precise
 Catalyzed by
spliceosome from snRPs
(small nuclear
ribonucleoproteins)
 Some RNA introns are
self splicing (ribozymes)
Alternative Splicing
 Process which allows
the production of a
variety of different
proteins from one
gene only
Genetic Code
 Set of rules specifying the relationship
between the sequence of bases in a DNA
or mRNA molecule and the order of amino
acids in the polypeptide chain encoded by
that DNA or mRNA
 Triplet code
Characteristics of Genetic Code
 Degenerate
- More than one nucleotide triplet can
specify one amino acid
 Specificity
- Specific codon  same amino acid
Gene expression lies in the relationship between the nucleotide base
sequence of DNA molecules and the linear order of amino acids in
protein molecules
Translation
 Production of a polypeptide by means of
an mRNA template
 Conversion of the information in the
nucleic acid sequence to polypeptides of a
specific amino acid sequence.
 Occurs in the cytoplasm
tRNA

Codon – triplet of
nucleotides in mRNA
Anticodon – triplet of
nucleotides in tRNA
Stages of Translation
 Initiation
 Elongation
 Termination
Initiation
 AUG  start of protein synthesis
 Initiation factors
 Initiator tRNA binds to the ribosome
Prokaryotes vs. Eukaryotes
 50S and 30S  60S and 40S
ribosomal subunit ribosomal subunit
 Shine Dalgarno  Kozak sequence
Sequence (AGGA) (A/GCCACCAUGG )

Purine

Note: RBS
Ribosomal
Binding Site
 A site – aminoacyl site
P site – peptidyl site
E site – exit site

70S Initiation Complex (Prokaryotes)/ 80S Initiation

Complex (Eukaryotes)
Protein synthesis: Translation initiation
Elongation
 Successive addition of amino acids in a
sequence specified by codons in mRNA
 Elongation factors (EF-Tu and EF-Ts)
 Steps: binding of aminoacyl tRNA, peptide
bond formation, translocation
 1. Aminoacyl
tRNA binding

3. Translocation
2. Peptide bond
formation
C-terminus N-terminus end
end

Peptide bond formation and

Translocation
 Protein
synthesis:
elongation (2)

http://www.phschool.com/science/biology_
place/biocoach/translation/term.html
Termination
 Stop codons (UAG, UAA and UGA)
 Release factors recognize stop codons
(mimic the appearance of tRNA)
Protein
translation:
termination (3)
Protein Translation: Summary
Overall Summary
Transcription
mRNA rRNA tRNA

Translation
Mutation
 Permanent change in DNA sequence of
the gene
What Causes DNA Mutations?
 Mistakes that occur when a cell copies its
DNA in preparation for cell division
 Environmental agents (UV light, X-ray,
intercalating agents)
Effects of Environmental agents
Effects of Environmental agents
Errors during DNA Replication
Type of Mutations
 Point Mutation
- Silent
- Missense
- Nonsense
 Frameshift Mutation
- Insertion
- Deletion
 mRNA

parental DNA
arginine glycine tyrosine tryptophan asparagine amino acids

altered mRNA

DNA with
base insertion
arginine glycine leucine leucine glutamate
altered amino-
acid sequence
A frameshift mutation changes the amino acid
sequence from the site of the mutation.
Sickle Cell Anemia
 Caused by point mutation
 Transposons
“Jumping Genes”
 Dr. Barbara McClintock
 Nobel Prize in Medicine in 1983
 Corn research (Indian corn)
 DNA segments that move spontaneously about
the genome
 When they insert into a gene region, they
usually inactivate that gene
 Purple pigment = “turned off”
 Transposons
“Jumping Genes”
= transposon

= purple pigmentation gene

 transposon adjacent  blocks

pigmentation gene is blocked  white area

• transposon  gene for pigmenttation gene

 production of purple pigment is resumed