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III. Translational Regulation
•NOTE: Difference between prokaryotic and eukaryotic
translation
Prokaryote Eukaryote
•Formylmethionine as first •Methionine as first amino
amino acid acid
•Shine-Dalgarno sequence •7mGppp – where rRNA
– where 16S rRNA binds binds
AGGAGG 10 bases
upstream of AUG codon
III. Translational Regulation
•Achieved by altering the half-life or stability of the mRNA for
translation
•e.g. removal of poly-A binding protein (PABP) reduces the
half-life of mature mRNAs.
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III. Translational Regulation
•Also achieved by controlling the initiation and rate of
translation
•e.g. iron-responsive element (IRE) of the 5’ UTR of human
ferritin gene.
Abundance of iron
5’ AUG Ferritin mRNA Ferritin Iron storage
Depletion of iron
5’ AUG Ferritin mRNA Ferritin Iron storage
= IRE = iron regulatory protein
IV. Post-translational Regulation
•Modification of protein product
•Ubiquitination – attachment of protein ubiquitin to other
proteins
– protein becomes target of destruction
by proteosome
•Phosphorylation – carried out by kinases
– addition of phosphate group to amino
acids
– affects activity of protein
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IV. Post-translational Regulation
•Modification of protein product
•Glycosylation – addition of carbohydrate group
– also affects protein activity
•Enzymatic cleavage – breakdown of protein into smaller
functional units
– carried out by peptidase
Remember: Structure of a prokaryotic gene
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Remember: Structure of a eukaryotic gene
EUKARYOTIC DNA
1. Nuclear DNA Arranged into chromosomes
2. Mitochondrial DNA
Circular DNA
3. Chloroplast DNA
•“extrachromosomal” or
“cytoplasmic” DNA
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•Chloroplast and mitochondria are semi-autonomous
•They have their own genes
•They can replicate themselves
Evidence for endosymbiotic theory of the origin
of these organelles
•Chloroplasts and mitochondria were once
prokaryotes that entered eukaryotic cells
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•There is interaction between nuclear and cytoplasmic DNA
•e.g. Ribulose-1,5-bisphosphate carboxylase/oxygenase
(RuBisCO)
•Probably the most abundant and most important
protein on earth
•Catalyzes first step in carbon fixation during the
Calvin cycle of photosynthesis
•There is interaction between nuclear and cytoplasmic DNA
CHLOROPLAST rbcL mRNA Large subunit
RuBisCO
NUCLEUS rbcs mRNA Small subunit
PLANT CELL
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DEVELOPMENTAL GENETICS
Differential Gene Expression
•Genetic basis of development
•All cells in an organism are GENOMICALLY EQUIVALENT
•Cells become structurally, functionally, & biochemically
different by expressing different genes at different times
during development
NOTE: Totipotent cells can give rise to an entire adult plant
(F.C Stewart’s experiment at Cornell University)
Culture medium provided sequentially
with cytokinins (for shoot formation)
and auxins (root formation)
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Cloning of Dolly
Differentiation of stem cells
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Northern Blot Analysis
•Genes are not necessarily
expressed in the same
organs.
Establishment of Polarity in Animals
•Genes determine polarity
•Genes yield transcription factors that signal other
genes to produce other transcription factors to signal
another set of genes cascade of gene switching
•Examples
•Maternal effect genes
•Zygotic genes
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Hox Genes
•Regulate development
•Determine which body parts are made at which locations on
developing organisms
•Encode transcription factors which typically switch on
cascades of other genes
•First found in the fruit fly Drosophila melanogaster
• Each includes a 180-bp domain known as homeobox
(encodes a DNA-binding sequence of 60 amino acids
known as homeodomain)
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• Homeobox-
containing genes
• Drosophila
• 1 chromosome
• Mammals
• 4 chromosomes
Life Cycle of Drosophila
melanogaster
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Mutations in Hox Genes
Mutation in Bithorax and Postbithorax genes
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Mutations in Hox Genes
Mutation in Antennapedia gene
(a) Head from wild-type Drosophila
(b) Head from an Antp mutant, showing the replacement of normal
antenna structures with legs – caused by activation of the Antp
gene in the head origin.
DNA Microarray Assays
•Automated detection and measure of expression of
thousands of genes at a given time
•Makes use of cDNAs amplified from mRNA
transcripts
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DNA Microarray Assays
• Spots where any of the cDNA hybridizes fluoresce with an
intensity indicating the relative amount of the mRNA that
was in the tissue.
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DNA Microarray Assays
•Information from this assay can provide:
•how ensembles of genes interact to form a living
organism.
•comparisons between cancerous and non-
cancerous cells.
GENETICS OF VIRUSES AND BACTERIA
•Bacteria and viruses are model genetic organisms
•small
•simple
•genes can be
manipulated
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Viruses
•Aggregates of nucleic acids and proteins
•Not considered living-----WHY?
•Types of nucleic acids used
•ds DNA
•ss DNA
•ds RNA
•ss RNA
Capsid – protein shell of
viral genome
Capsomere – building block
of capsid
Viruses
Viral envelope – membrane
covering capsids
– derived from host
cell
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Viruses
•Bacteriophage (phage)
– virus that infects bacteria
Viruses
•Viruses can reproduce only within
a host cell
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Viruses
•Mechanisms of viral infection
1. Lytic cycle – results in lysis (death) of host cell
Viruses
•Some bacterial defenses against lytic phages:
•Change in receptor sites for phages
•Restriction nucleases cut foreign DNA
•BUT natural selection favors resistant phages
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Viruses
•Mechanisms of viral infection
2. Lysogenic cycle – no lysis of host cell
NOTE: Temperate phage – phages that use both cycles
•e.g. lambda phage that infects E. coli
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Viruses
•Retroviruses – contain reverse transcriptase
enzyme
– e.g. Human immunodefficiency virus (HIV)
Viruses
•Retroviruses
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Viruses
•Probable origins
•Probably evolved after the first cells
•Hypothesis: originated from fragments of cellular nucleic
acids that could move from one cell to another
•Plasmid – small, circular DNA in bacteria and yeast
– replicate independently
•Transposon – DNA segments that move from one
location to another within a cell’s
genome
Other Replicating Particles
•Viroids – naked circular RNAs that infect plants
– do NOT code for proteins
– disrupt plant growth and other metabolic
processes
Potato spindle tuber viroid
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Other Replicating Particles
•Prions – replicating infectious PROTEINS
– may cause neurological disorders i.e. mad cow
disease and Creutzfeldt-Jacob in humans
Other Replicating Particles
•Prions – replicating infectious PROTEINS
•Kuru disease – an incurable degenerative neurological
disorder leading to spongiform encephalopathy and
caused by a prion found in humans
•was transmitted among members of the Fore
tribe of Papua New Guinea via cannibalism
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