An Introduction of Gene
Function
Gene participates in three major
activities:
A. A gene is repository of information
Transcription Translation
DNA RNA Protein
B. A gene can be replicated
DNA
replication
C. A gene can accept occasional changes,
or mutation
Signals DNA Recombination
(Stimulate) Mutation
The Central Dogma Outlines the Flow of
Genetic Information
Outline of gene expression
Gene expression: the process by which gene products
are made.
(Sense strand)
(Antisense strand)
Transcription
Initiation, elongation and termination
The enzyme recognizes a region calls a promoter,
which lies just “upstream” of the gene.
The polymerase, RNA polymerase, binds tightly to
promoter and causes localized melting, or
separation , of the two DNA strands within the
promoter.
The RNA polymerase starts building the RNA chain.
The substrates are the four ribonucleoside
triphosphates.
The first substrate is usually a purine nucleotide.
After the first nucleotide is in place,
the polymerase joins a second nucleotide
to the first, forming the initial
phosphodiester bond in the RNA chain.
During the elongation phase of
transcription, RNA polymerase directs the
sequential binding of ribonucleotides to the
growing RNA chain in 5’ 3’ direction.
The region of the ends of genes, called
terminator, signal termination. These work
in conjunction with RNA polymerase to
loosen the association RNA product and
DNA template.
Making RNA
Various Classes of RNA Have
Different Functions
Problem
The following is the sequence of the template strand
of a DNA fragment:
5’ ATGCGTGACTAATTCG 3’
a. Write the sequence of this DNA in conventional
double-stranded form.
b. Assuming that transcription of this DNA begins
with the first nucleotide and ends, write the
sequence of the transcript of this DNA in
conventional form (5’ to 3’ left to right)
Patterns of transcription
Translation
There are two substances play key roles in
translation: ribosome and transfer RNA
Ribosome: Proteins-synthesizing machines
Transfer RNA: The adapter molecule
Ribosomes Read the Genetic Code
Ultracentrifugation
(a very high-speed centrifuged)
• It is able to generate about 700,000 folds gravity.
• The molecule particles sediment speed dependent on two
properties in ultracentrifugation.
– molecular weight: Heavy particles sediment more rapidly
than light ones.
– molecular shape: spherical particles migrate faster than
extended or flattened ones.
• Sedimentation coefficient:
– S = velocity/ centrifuge force = v/w2r,
– v= w2rm (1-VB)/f
– 1s = 10 -13 sec
• W= 角速度, m= mass, B= solvent density, f= 摩擦係數
• S depend on molecular weight and shape.
Sucrose density gradient
centrifugation
(蔗糖密度梯度離心)
S: Sedimentation coefficient (沈降係數）
Ribosome : Prokaryotic cells (70S ribosome = 30S + 50S)
30S ribosome (21 proteins + 16S ribosomal RNA)
50S ribosome (36 proteins + 23S and 5S ribosomal RNA)
Eukaryotic cells (80S ribosome = 40S + 60S)
40S ribosome (33 proteins + 18S ribosomal RNA)
60S ribosome (49 proteins + 28S、5.8S and 5S ribosomal RNA)
Transfer RNA: The adapter molecule
The tRNA has two
“ business end”. One end
attaches to an amino acid. The
other end contains a 3-bp
sequence that pairs with a
complementary 3-bp sequence
in an mRNA.
The enzyme call aminoacyl-
tRNA synthetase that play a
specific amino acid attach to a
specific tRNA. For example,
phenylalanine –tRNA
synthetase catalyze
phenylalanine link to
phenylanine-tRNA.
Codon-anticodon recognition
The Genetic Code Dictates the Amino Acid
Sequence of Proteins
Translation initiation
The initiation codon : AUG
The initiation codon interacts with a special
aminoacyl-tRNA. In eukaryote this is methionyl-
tRNA ( a tRNA with methionine attached); in
prokaryotes it is a derivative called N-
formylmethionyl-tRNA.
The prokaryotic messages have a special
sequence, called a Shine-Dalgarno sequence,
just upstream of the initiating AUG. This
sequence attracts ribosomes to the nearby
AUG so translation can begin.
Eukaryotic mRNAs have a special
methylated nucleotide called a cap at their
5’end. A protein called the cap-binding
protein (CBP) binds to the cap and then helps
attract ribosome.
Translation elongation
Translation termination
Termination codon (stop codon): UAG, UAA
and UGA
Protein factors: release factors
The release factors recognize these stop
codons and cause translation to stop, with
release of the polypeptide chain.
Simplified gene and mRNA structure
fMet-His-Gly-Ile
Proteins Carry Out Many
Cell Functions
• Proteins are made from a linear chain
of monomers, known as amino acids, and
are folded into a variety of complex
3D shapes.
• A chain of amino acids is called a
polypeptide chain.
The amino acids join together in proteins
via peptide bonds.
Proteins contain 20 different amino acids. Each amino
acids has an amino group (NH3+), a carboxyl group (COO-
), a hydrogen (H), and a side chain.
A polypeptide has a free amino group at its left end.
This is the amino terminus, or N-terminus. It also has a
free carbon group at its right end, which is the carboxyl
terminus, or C-terminus.
Protein function
• Some proteins provide the structure that
helps give cells integrity and shape, e.g.
actin.
• Some proteins serve as hormones to
carry signals from one cell to another,
e.g. insulin.
• Proteins can also bind and carry
substances, e.g. hemoglobin.
• Proteins also control the activities
of gene.
• Proteins serve as enzymes that
catalyze the hundreds of chemical
reactions necessary for lift.
Three hypothesis for DNA replication
Mutation
Mutation is essential for evolution.
Harmless mutation:
Silent mutation: a nucleotide
change not affect the protein.
AAAAAG
Lys Lys
Conservation change in amino acid,
e.g. a leucine changed to an
isoleucine.
The new amino acid is much different
from the old one, the change
frequently impairs or destroys the
function of the protein.
Sickle cell disease
What causes this sickling of red
blood cells?
This problem is in hemoglobin.
What is the difference between
normal hemoglobin (HbA) and sickle
cell hemoglobin (HbS)?
The sickle cell mutation and its
consequences
GAG GTG
Fingerprinting a protein
Sequences of amino-terminal peptides
from normal and sickle cell -globin
