Gene Expression
Gene Expression
Jularbal
DNA Structure
DEOXYRIBUNUCLEIC ACID
• Polymer of DEOXYRIBONUCLEOSIDE
MONOPHOSPHATES covalently linked by
3'→5'–PHOSPHODIESTER BONDS.
• DNA exists as a double- stranded molecule,
two strands forms a DOUBLE HELIX.
• DNA is found associated with nucleoprotein
present in the nucleus.
CHEMICAL COMPONENTS
DNA POLYMERASE
SEMICONSERVATIVE
PRIMASE
• Synthesize RNA primers
HELICASES
• Unwind the helix.
SSBP
• Hold the single-stranded conformation.
TOPOISOMERASES
• Prevents the extreme supercoiling of the parental
helix that would result because of unwinding at a
replication fork.
• Topoisomerases break and rejoin DNA chains.
• DNA GYRASE, a topoisomerase inhibited by the
quinolone family of antibiotics, is found only in
prokaryotes.
PRIMING
DNA POLYMERASES
PROKARYOTES
• Have three DNA polymerases: pol I, pol II, and pol III.
EUKARYOTES
RNA PRIMER
LEADING STRAND
• Formed by continuous copying of the parental
strand that runs 3′ to 5′ toward the replication
fork.
LAGGING STRAND
• Formed by discontinuous copying of the
parental strand that runs 3′ to 5′ away from the
replication fork.
• As more of the helix is unwound, the synthesis of
the lagging strand begins from another primer
forming short fragments known as OKAZAKI
FRAGMENTS.
• The RNA primers are removed by
NUCLEASES then the resulting gaps are filled
with the appropriate deoxyribonucleotides by
another DNA polymerase.
• Okazaki fragments are joined by DNA
LIGASE, an enzyme that catalyzes the formation
of phosphodiester bonds between two
polynucleotide chains.
RNA Structure
RNA
• Similar to DNA except that RNA
contains RIBOSE and URACIL.
• Single-stranded and can act as
CATALYSTS of reactions
• RIBOZYMES- usually precursors of
rRNA, remove internal segments of
themselves, splicing the ends together.
• RIBONUCLEASES- cleaving other
RNA molecules
• PEPTIDYLTRANSFERASE- an
enzyme in protein synthesis.
MESSENGER RNA (mRNA)
• Provide the blueprint for an amino acid
sequence.
• mRNA is about 5% of total RNA.
PROMOTER REGIONS
• Unwinds the DNA helix and transcribes the DNA template strand into RNA,
beginning at the transcription start site.
• The DNA strand complementary to the template strand is the CODING, or
SENSE STRAND.
• The RNA chain is elongated in the 5’ to 3’ direction.
• RIBONUCLEOSIDE TRIPHOSPHATES are added to the free 3’-OH
end of the RNA, producing a new 3’-OH terminal.
ELONGATION
TERMINATION
PROCESSING
INITIATION
INITIATION
• DNA unwinds and separates to form a
small open complex.
• RNA polymerase binds to the DNA of the
gene at a region called the PROMOTER
• INITIATION COMPLEX, containing
RNA polymerase II and transcription factors
is assembled near the Promoter region.
TATA BOX
• A conserved promoter sequence about 25
bases upstream of the start site.
• Helps orient the RNA polymerase.
CAAT BOX
• Conserved sequence within the promoter
region about 70 bases upstream of the start
site.
CG BOX
• Binds a specific TRANSCRIPTION
FACTOR (SP-1)
ELONGATION
ELONGATION
EXONS
INTRONS
RHO-DEPENDENT
TERMINATION
POLYADENYLATION
• Polyadenylation at the POLY(A) SITE leads to formation
of the POLY(A) TAIL consisting of ADENYLATE
RESIDUES at the 3’ end of the transcript.
SPLICING
SPLICE SITES
• POSITIVE CONTROL-factors
required to initiate RNA synthesis.
• NEGATIVE CONTROL - factors
required to prevent initiation of RNA
synthesis.
EUKARYOTIC GENE
EXPRESSION
• Regulated by controlling physical access
to the DNA by the RNA polymerase
and by controlling the rate of its
binding.
LAC OPERON SYSTEM
NEGATIVE CONTROL
• Occurs when only glucose is available as an
energy source and expression of the lac operon
is not needed.
• REPRESSOR PROTEIN is needed to inhibit
transcription
• LAC REPRESSOR PROTEIN is produced
constitutively and binds to the operon to
prevent transcription.
POSITIVE CONTROL
• Occurs when lactose is present, and expression
of the lac operon is needed.
• ACTIVATOR PROTEIN is needed to start
transcription
LACTOSE IS PRESENT
NO GLUCOSE IS PRESENT
GLUCOSE IS PRESENT