INTRODUCTION TO

CELL BIOLOGY

Dr Jeff O’Sullivan
Associate Professor in Biological sciences
School of Dental Science
Trinity College Dublin

jeff.osullivan@dental.tcd.ie
COURSE OUTLINE
Evolution/Cellular Evolution
Cellular Chemistry & Biosynthesis
Cell Structure and Function
Cell Metabolism
Cellular Replication
Cells: From DNA to Protein

Cell Cycle
Mutations
Cancer
Consequences
The three major divisions (domains) of the living world
 Evolutionary processes

8-6 X 10 years
7 2X 10 years
5
Evolutionary processes
Times of divergence of different
vertebrates
Evolutionary processes
 Miller-Urey Experiment 1953

"A Production of Amino Acids Under Possible Primitive Earth Conditions". Science 117. pp. 528–529
Prokaryote Eukaryote

0.5-100um 10-150um
The major features of eukaryotic cells
Elements in red account for ~99% of atoms in a human body
Elements in blue account for ~0.9% of atoms in a human body
Elements in green represent trace amounts
Elements in yellow????
Water (H2O) is the most abundant molecule in the body
H2O molecules are held together by hydrogen bonds

Water soluble molecules = Hydrophilic

Water insoluble = Hydrophobic
 Cells Contain Four Major Families
of Small Organic Molecules

Macromolecules (Big) are important chemical

components of the cell
These macromolecules are polymers made up of building
blocks (monomers)
 S-, N-, C-, and O-glycosidic bonds

Sugars → Polysaccharides → Signalling/Energy

Fatty Acids → Lipids

Triacylglycerol (Energy Storage)

Phospholipids (Membranes)
Amino Acids → Protein
FR7126
 Protein folds into precise 3-dimensional structure
that confers specific activity on protein e.g. enzyme
 Protein 3D structure

HIV-RT GFP Myoglobin

Nucleotides Nucleic Acids (DNA / RNA) Inheritance
 DNA Genes Chromosomes
Genes differ in their nucleotide sequence
(Enormous Variety Possible (4n))

Base pairing rules

DNA is a double helix

Each DNA turn is made up
of 10.4 nucleotide pairs
DNA replication is necessary prior to cell division
DNA replication is semi - conservative
DNA synthesis catalysed by DNA polymerase
DNA synthesis takes place in 5’ to 3’ direction
 DNA replication:
Synthesis of one stand is continuous
Synthesis of other strand is discontinuous
Replication
overview
 DNA encodes for Proteins

The unit of DNA that encodes for a protein is a GENE

We have ~ 25,000 genes

Genes are linked together to form CHROMOSOMES

Chromosomes consist of DNA that winds around proteins (Histones)

DNA + Histones = Chromatin

Macroscopic chromosome structure
 Nucleosomes are a basic unit of
eukaryotic chromosome structure
 From DNA to Protein
Two stage process
1: DNA RNA (Transcription)
2:RNA Protein(Translation)
Ribonucleic Acid
(RNA)
 Transcription

Transcription is catalysed by RNA

polymerase II (Pol II)

Transcription is initiated by binding

of Pol II to start site of gene
(at a region known as the
promoter)

Binding is regulated by proteins

called transcription factors

Pol II makes a RNA strand

(messenger RNA (mRNA))
complementary to one of DNA
strands
Transcription

Initiation of transcription by RNA

polymerase II in eukaryotic cells
The pre-mRNA
splicing reaction
Nucleotide sequence of gene determines amino acid sequence of protein
DNA initially copied into RNA by DNA transcription

Messenger RNA (mRNA); Transcripts that direct synthesis of protein

Other RNA transcripts: Transfer RNA (tRNA) and ribosomal RNA (rRNA)

mRNA initially produced as primary transcripts containing intron and exon sequences

Introns removed by RNA splicing in nucleus (Increases variety of proteins)

Nucleotides in mRNA are “read” in serial order in sets of 3 (codons)

Codon specifies amino acid

Genetic code made up of 64 codons

Genetic code is degenerate

3 possible reading frames.

Only 1 is used

mRNA is read by
Ribosomes
Ribosomes are made up of proteins and ribosomal RNA (rRNA)

Prokaryotic and Eukaryotic Ribsomes differ in their composition

Some useful antibiotics specifically inhibit prokaryotic ribosomes
and so selectively target bacterial protein synthesis
eg. Tetracycline, Streptomycin, Chloramphenicol, Erythromycin
 Translation and transfer RNA (tRNA)
Ribosomes read mRNA from 5’ to 3’ (codon by codon)

tRNA acts as adaptor molecules and match amino acids to

codons via anticodons

Aminoacyl-tRNA linkage catalysed by specific aminoacyl-tRNA synthetases

(Proof-readers)

Wobble base-pairing allows for one tRNA to recognise codons that differ in
their last nucleotide (→ degeneracy)
Initiation of Translation
Catalysed by initiation factors (IFs)
which bind to small ribosomal subunit

Initiator tRNA (for methionine) loaded

onto small ribosomal subunit by
eukaryotic initiation factor 2 (elF-2)

Loaded small ribosomal subunits binds to

mRNA (via 5’cap) and scans for first
start codon (AUG) (Methionine)

Initiation factors dissociate and large

ribosomal subunit binds

Initiator tRNA now bound to P site

(peptidyl-tRNA-binding site)

Second aminoacyl-tRNA (with associated

elongation factor [EF]) binds to A site
(aminoacyl-tRNA-binding site) via codon-
anti-codon interactions
 Amino acid in P site uncoupled from
tRNA and joined by peptide bond to
amino acid in A site
(catalysed by peptidyl transferase)

New peptidyl-tRNA in A site translocated

to P site by ribosome moving exactly
three nucleotides along mRNA

Free A site occupied by new aminoacyl-

tRNA with appropriate anti-codon

Carboxyl end of peptide in P site forms

peptide bond to amino acid in A site etc

Ribosome reaches stop codon (UAA, UAG

or UGA)

Release factors bind and polypeptide

released by hydrolysis

Ribosome releases mRNA and dissociates

for re-initiation of translation

Multiple ribosomes can simultaneously

translate the same mRNA (Polysomes)
Overview of Process of Protein Production
PRESENTATION: PROTEIN
• Select a protein of your choice

• 3-5 min presentation describing the characteristics

• PowerPoint

• Send slides by Mon 9th Oct

 The Cell Cycle
Cell reproduction

Specific sequence of orderly events

Results in:

Two daughter cells

Duplication of contents

Primarily replication of DNA

Accurate distribution between

daughter cells
 The Cell Cycle
Four phases

S phase (10-12h)
DNA synthesis

M phase (<1h)
Mitosis….no cell growth
G1 phase
Gap/growth
G2 phase
Gap/growth

G0 phase
Resting
The Cell Cycle

DNA content during cell cycle

unreplicated DNA…..G1
fully replicated DNA…..G2/M
intermediate amount of DNA…..S
Control of The Cell Cycle

Mediated by CDKs
(Cyclin-Dependent Protein Kinases)
 Cyclin-Dependent Protein Kinases
G1/S-cyclins
Activate CDKs in late G1…commit to cell-cycle entry

S-cyclins
Bind CDKs after start and stimulate chromosome duplication

M-cyclins
Activate CDKS that stimulate entry into mitosis at G2/M checkpoint

G1-cyclins
Govern activities of G1/S-cyclins controlling progression through Start in late G1
Cyclin-Dependent Protein Kinases
 Mutations: Effect Protein Function

Antigenic Drift/Shift Silent/Non-Silent

Good and Bad

Evolution Dysfunction

Sickle Cell Anaemia Cancer

HALLMARKS OF CANCER
Carcinogenesis or Oncogenesis or Tumorigenesis:
The creation of cancer
Cancer cells reproduce without restraint and colonise other tissues

Carcinomas Epithelial cells

Sarcomas Connective tissue/Muscle cells
Leukemias/Lymphoma White blood cells
 Most cancers derive from a single abnormal cell

Single mutations are

not enough

Requires substantial
number
of rare genetic
accidents
Cancers develop gradually from increasingly aberrant cells
 To metastaize cells must proliferate
and survive in a foreign environment

Tumors induce angiogenesis

Hypoxia Angiogenic Switch HIF1a VEGF Blood vessels
 Properties of Cancerous Cells/Growth
More self-sufficient then normal cells

Insensitive to anti-proliferation signals

Less prone to apoptosis

Defective in intracellular control mechanisms e.g. stress

Induce help from normal stromal cells

Induce angiogenesis

Metastasis

Genetically unstable

Can stabilize telomeres

Overcoming cancer
Overcoming cancer
Detecting cancer
Understanding cancer