Gene Expression

INTRODUCTION
An essential characteristic of all living
organisms is their ability to store and express
genetic information.
The genetic information in a human cell is
contained within several DNA molecules.
DNA contains both unique coding sequences
(EXONS) that are expressed and noncoding
sequences (INTRONS) that are not expressed
 INTRODUCTION
In the human genome, only about 1.5% of the
DNA is functional.
During cell division, DNA molecules are
duplicated via a process called replication and
then passed on to each daughter cell.
 INTRODUCTION
 Within a cell, the information in DNA is expressed
through the formation of RNAs and proteins.
 Transcription involves making RNA molecules that are
complementary to one of the two strands of DNA.
 There are 3 main types of RNA: messenger RNA
(mRNA), transfer RNA (tRNA) and ribosomal RNA
(rRNA).
 Transcription involves enzymes called RNA
polymerases that bind to the DNA and separate the 2
strands to create a transcription bubble
 INTRODUCTION
 As transcription occurs, ribonucleotides are
joined together as follows: A pairs with T, U
pairs with A, G pairs with C, and C pairs with
G. As the RNA polymerase moves along the
DNA, a new RNA strand is formed in the 5’ 3’
direction.
 Transcription begins at specific sites within
the DNA called promoters and ends at sites
within the DNA called terminators.
 INTRODUCTION
 Translation involves using messenger RNAs to direct the
formation of specific proteins by the ribosomes (protein
factories).
 The mRNA is read in 3-letter blocks or codons.
 The genetic code is unambiguous in that each codon has
one specific meaning.
 The code is degenerate in that some amino acids are
encoded for by more than 1 codon.
 The codon AUG, which specifies methionine, is used to
START translation.
 The codons UAA, UAG, and UGA do not encode for amino
acids and are used to STOP translation.
 INTRODUCTION
 Transfer RNAs are RNA molecules that transfer amino acids to
the growing polypeptide chain.
 tRNAs have a characteristic clover-leaf shape and an anticodon
region that is complementary to the codon region of the
mRNA.
 Amino acids are attached to tRNA by enzymes called
aminoacyl-tRNA synthetases.
 Ribosomes are RNA/protein complexes composed of large and
small subunits.
 The complete ribosome binds to mRNA and has 3 sites at
which tRNAs can bind: the aminoacyl site (A) site, the peptidyl
(P) site, and the exit (E) site.
A
 INTRODUCTION
 During the initiation phase of translation, a
mRNA combines with the small and large
ribosomal subunits and a transfer RNA carrying
methionine binds to the start codon (AUG).
 During the elongation phase of translation,
several tRNAs carrying amino acids bind to the
ribosome as it moves along the mRNA and the
amino acids are linked together by peptide
bonds.
 During the termination phase of translation,
the ribosomal subunits release both the
mRNA and the newly formed protein.
 These proteins then carry out specific
functions to give each cell its characteristic
properties.
 While replication and transcription of DNA
occur in the nucleus, translation occurs in the
cytoplasm.
Before making proteins, Your cell must first
make RNA
 Question:
 How does RNA (ribonucleic acid) differ from
DNA (deoxyribonucleic acid)?
 RNA differs from DNA
1.RNA has a sugar ribose
DNA has a sugar deoxyribose

2. RNA contains uracil (U)

DNA has thymine (T)

3. RNA molecule is single-stranded

DNA is double-stranded
Gene expression
 Gene Expression
Replication: DNA---- DNA copy
(occurs in the nucleus).
 Transcriptation: DNA -----mRNA
( Occurs in nucleus)
Translation: mRNA --- protein
(occurs in the cytoplasm)
 Type of RNA Function in Function

Messenger Nucleus, migrates Carries message from DNA to ribosome in the

RNA(mRNA) to ribosomes cytoplasm
in cytoplasm

Transfer RNA (tRNA) Cytoplasm Transfer amino acid to ribosome durning

protein synthesis

Ribosomal Cytoplasm (rRNA)found in ribosome, joins mRNA and

RNA(rRNA) tRNA; forms protein

snRNA Small nuclear RNA, forms complexes with proteins used

in eukaryotic RNA processing (e.g., exon splicing and
intron removal).

Micro RNA Micro RNA/small interfering RNA, short ~22 nt RNA

sequences that bind to 3’ UTR target mRNAs and result in
gene silencing.
Codon= sequences that codes for a protein
Anticodon= code complementary to mRNA
 Transcriptation
Transcriptation=DNA mRNA
Promoter: DNA site where transcription stars
RNA polymearse adds A,U,C,G
Transcription is a process in which one DNA strand is
used as template to synthesize a complementary RNA
Note that
uracil (U) of RNA is paired with adenine (A) of DNA.
There are a few different names for these nucleic acid
strands.
The DNA strand which serves as the template may be
called "template strand", "minus strand", or "antisense
strand".
The other DNA strand may be termed "non-template
strand", "coding strand", "plus strand", or "sense strand".
Since both DNA coding strand and RNA strand are
complementary to the template strand, they have the
same sequences except that T in the DNA coding strand
is replaced by U in the RNA strand.
 Transfer RNA (tRNA)
amino acid
attachment site methionine amino acid

U A C
anticodon
 How your cell makes very important
proteins
 The production (synthesis) of proteins.
 3 phases:
1. Transcription
2. RNA processing
3. Translation
 DNA  RNA  Protein
 Transcription OR RNA production

RNA molecules are produced by copying part of

DNA into a complementary sequence of RNA
This process is started and controlled by an
enzyme called RNA polymerase.
Only one strand of DNA is copied.
A single gene may be transcribed thousands of
times.
After transcription, the DNA strands rejoin
 Steps involved in transcription

The enzyme RNA polymerase is responsible for

creating RNA by copying the template strand of
DNA.
Before transcription can begin in eukaryotes,
proteins called transcription factors must bind to
a region of the DNA called the promoter.
The promoter identifies the start of a gene, which
strand is to be copied, and the direction that it is
to be copied.
RNA polymerase binds to the transcription factors
and the promoter
 2. RNA Processing

 Introns are pulled out and exons come together.

 End product is a mature RNA molecule that
leaves the nucleus to the cytoplasm.

 Introns bad…… Exons good!

 2. RNA Processing
pre-RNA molecule

exon intron exon intron exon

intron intron

exon exon exon

splicesome splicesome

exon exon exon

Mature RNA molecule
Post-transcriptation modification
 Post-transcriptation modification
A modified guanine nucleotide "cap" is added
to the 5’ end and a poly-A tail (50 to 250
adenines) is added to the 3’end of the
molecule.
These modifications are thought to:
1) enhance the movement of mRNA through the
nuclear pores into the cytoplasm
2) prevent the destruction of mRNA by hydrolytic
enzymes,
3) and help ribosomes attach during translation
Noncoding segments,
introns, are spliced out DNA
Exon Intron Exon Intron Exon

A cap and a tail are Cap

Transcription
Addition of cap and tail

added to the ends RNA

transcript
with cap Introns removed
and tail

Exons spliced together

mRNA

Coding sequence
NUCLEUS

CYTOPLASM
The 5' end and the 3' end each contain
nucleotides that are not translated into
protein.
These two regions are called the 5' UTR
(untranslated region) and the 3' UTR.
 Production of the mRNA molecule

Three main parts:

1. 5’ untranslated region (5’ UTR) or leader sequence

2. Coding sequence, specifies amino acids to be translated

3. 3’ untranslated region ( 3’ UTR) or trailer sequence

may contain information that signals the stability of the particular mRNA
 Splicing

 Eukaryotic genes contain regions that are not

translated into proteins.
 These regions of DNA are called introns (intervening
sequences) and must be removed from mRNA by a
process called RNA splicing.
 Their function is not well understood.
 The remaining portions of DNA that are translated
into protein are called exons (expressed).
 After intron-derived regions are removed from mRNA,
the remaining fragments- derived from exons- are
spliced together to form a mature mRNA transcript.
 Exon Intron Exon Intron Exon
DNA
• Noncoding segments, Transcription
Addition of cap and tail

introns, are spliced RNA

transcript

out with cap

and tail

• A cap and a tail are

Exons spliced together

added to the ends mRNA

Coding sequence
NUCLEUS

CYTOPLASM
 Next amino acid
tRNA Growing to be added to
molecules polypeptide polypeptide
P site A site
Growing
Large polypeptide
subunit
tRNA

P A
mRNA
mRNA
binding
site
Codons

mRNA Small
subunit
 3. Translation
 Three parts:
1. initiation: start codon (AUG)
2. elongation:
3. termination: stop codon (UAG)
 Proteins
Proteins have a variety of roles that they must fulfil:
1. they are the enzymes that rearrange chemical
bonds.
2. they carry signals to and from the outside of the
cell, and within the cell.
3. they transport small molecules.
4. they form many of the cellular structures.
5. they regulate cell processes, turning them on
and off and controlling their rates.
 Proteins – Amino Acids
there are 20 different types of amino acids
different sequences of amino acids fold into different 3-D
shapes.
Proteins can range from fewer than 20 to more than 5000
amino acids in length.
Each protein that an organism can produce is encoded in
a piece of the DNA called a “gene”.
the single-celled bacterium E.coli has about 4300
different genes.
Humans are believed to have about 30,000 different genes
(the exact number as yet unresolved),
 Name 1-letter code Triplet Protein-Sequence
Glycine G GGT,GGC,GGA,GGG
(Alphabet:
Alanine A GCT,GCC,GCA,GCG
Valine V GTT,GTC,GTA,GTG
ACDEFGHIKLMNPQRSTVWY)
Leucine L TTG,TTA,CTT,CTC,CTA,CTG :
Isoleucine I ATT,ATC,ATA
Histidine H CAT,CAC
Serine S TCT,TCC,TCA,TCG,AGT,AGC
MENFQKVEKIGEGTYGVVY
Threonine T ACT,ACC,ACA,ACG KARNKLTGEVVALKKIRLDT
Cysteine C TGT,TGC ETEGVPSTAIREISLLK...
Methionine M ATG
Glutamic Acid E GAA,GAG
Aspartic Acid D GAT,GAC,AAT,AAC
Lysine
Arginine
K
R
AAA,AAG
CGT,CGC,CGA,CGG,AGA,AGG
• A typical human
Asparagine
Glutamine
N
Q
AAT,AAC
CAA,CAG
cell contains
Phenylalanine F TTT,TTC
about 100 million
Tyrosine Y TAT,TAC
Tryptophan W TGG proteins of about
10,000 types
Proline P CCT,CCC,CCA,CCG

Terminator (Stop) * TAA,TAG,TGA

RNA genetic code
Proteins
Primary protein structure
is the sequence of a chain of amino acids

Secondary protein structure

occurs when the sequence of
amino acids are linked by hydrogen
bonds.

Tertiary protein structure

occurs when certain attractions
are present between alpha helices
and pleated sheets.

Quaternary protein structure

is a protein consisting of more
than one amino acid chain.
 Proteins - Summary
DNA sequence determines protein sequence
Protein sequence determines protein
structure
Protein structure determines protein folding
and function