Chapter 15: Gene Expression
[Link]
Nov. 11: Reminders & Agenda
• Thursday Nov. 12 Ch. 15 Homework
• Friday Nov. 13 Key Assignment Meeting #3 Summary
• Friday Nov. 13: Last day to withdraw from the course with a
“W”
• Saturday Nov. 14 Lab Review 14: Natural Selection
• Tuesday Nov. 17 Ch. 16 Homework
• Thursday Nov. 19 Ch. 17 Homework
• Key Assignment Video Friday Nov. 20
• Tuesday Nov. 24 Exam 5 over Ch. 15, 16, 17
Part A: Gene Expression Objectives
• Understand the “Central Dogma” of molecular
biology
• Be familiar with transcription in prokaryotes
• Understand the role of promoters in
prokaryotic transcription
• Understand how transcription is terminated
• Be familiar with transcription in eukaryotes
• Understand how pre-mRNA is modified in
eukaryotes
Gene (DNA) to mRNA to Protein
• Remember: a
gene is a
molecular
blueprint to
synthesize a
protein.
Central Dogma: Information Flow
DNA RNA Protein
Transcription Translation
Information is The information The information in
stored in DNA. in DNA is copied RNA guides the
into RNA. production of proteins.
Gene (DNA) to mRNA to Protein
• How do you go
from DNA to
mRNA?
• What factors are
involved in this
process?
• Let’s brainstorm!
From Gene to mRNA to Protein
Inside nucleus TRANSCRIPTION Cytoplasm TRANSLATION
The gene’s sequence is copied The gene’s sequence is now
from DNA to a middleman encoded in mRNA, which directs
molecule called mRNA. the production of a protein.
mRNA
DNA
RNA polymerase
Genes Protein molecule
Ribosome
mRNA Nuclear
pore
Grandmother’s Copying cookie Index card Combining and Chocolate
cookbook recipe to index card with recipe baking ingredients chip cookies
• For more information see Slide 7
notes.
Flow of genetic
information
• Instructions on DNA are
transcribed onto
messenger RNA.
• Ribosomes are able to
read the genetic
information inscribed on a
strand of messenger RNA.
• They use this information
to string amino acids
together into a protein.
Take Home Message
q The genes in strands of DNA are a storehouse
of information, an instruction book.
q Theprocess by which the information is used
to build an organism occurs in two main steps:
q Transcription – in which a copy of a gene’s
base sequence is made
q Translation – in which that copy is used to
direct the production of a polypeptide.
In transcription, the information coded in DNA is
copied into mRNA.
• Information coded in DNA is transferred to a complementary RNA copy
• Similar to DNA replication, except:
1. Only one DNA strand used as a template
2. Only transcribes one gene at a time
3. RNA polymerases used to form RNA strands
4. Resulting RNA molecules are single strands
5. Uracil replaces thymine
In transcription, the information coded in
TRANSCRIPTION TRANSLATION
DNA is copied into mRNA.
DNA
1 RECOGNIZE AND BIND 2 TRANSCRIBE 3 TERMINATE
Once RNA polymerase As the DNA strand is processed through When the RNA polymerase
recognizes a promoter site, the RNA polymerase, the RNA polymerase encounters a code signaling
it binds to one strand of the builds a single- stranded RNA copy of the the end of the gene, it stops
DNA and begins reading gene, called the mRNA transcript. transcription and releases
the gene’s message. the mRNA transcript.
RNA RNA
polymerase RNA polymerase
polymerase
Promoter site Termination site
mRNA transcript
mRNA
transcript
UNWIND AND REWIND
As the RNA polymerase moves
down the strand of DNA, the
helix unwinds so that the DNA 4 CAP AND PROCESS
can be read. At the same time, In eukaryotes, mRNAs
the DNA that has already been receive extra processing
transcribed rewinds back to its Tail before they can be
original double-helix form. translated into a protein.
Cap A cap and tail are often
Helix unwinds Non-protein-coding
regions of mRNA added for protection and
to promote recognition,
and noncoding sections
are removed.
Helix rewinds
mRNA leaves the nucleus to be
translated into a protein.
• For more information see
Slide 11 notes.
Transcription: DNA to RNA
• No primers needed to start • Three key steps:
complementary copy 1. Initiation
2. Elongation
• RNA is made in the 5´→ 3´ 3. Termination
direction
Transcription Overview (1)
Transcription Overview (2)
Transcription Overview (3)
• RNA polymerase adds RNA nucleotides
sequentially according to the DNA template
Transcription Overview (4)
• RNA polymerase and completed RNA
transcript release from DNA template
Organization of a Gene (DNA)
• Promoter
– Control sequence initiates transcription
• Transcription unit
– Portion of gene that is copied into RNA
• Terminator
– Signals the end of transcription of a gene
Initiation at the Promoter
• In prokaryotes RNA polymerase is composed of 5 subunits: ⍵, ⍺, ⍺, β, β’, (and σ)
• ⍺ subunits: assemble RNA polymerase on DNA
• β subunits: bind to incoming RNA nucleotides (RNA subunits)
• σ subunit: initiation of transcription
• The σ subunit of prokaryotic RNA polymerase recognizes consensus sequences found in the
promoter region upstream of the transcription start sight. The σ subunit dissociates from the
polymerase after transcription has been initiated.
Transcription: Initiation
Transcription: Elongation
Elongation
• During elongation, the prokaryotic RNA polymerase tracks along the DNA
template.
• Then it synthesizes mRNA in the 5' to 3' direction and unwinds and rewinds the
DNA as it is read.
• Elongation occurs at a rate of 40 nucleotides per second.
Transcription: Termination
Termination
• Liberates the mRNA
strand
• Termination signal:
• Rho-dependent
(controlled by the
Rho protein)
Termination
• Liberates the mRNA
strand
• Termination signal:
• Rho-independent
(controlled by
sequences in the
DNA strand)
Transcription in Eukaryotes: Types of
eukaryotic polymerases
• RNA polymerase I
• Transcribes ribosomal RNA genes to make
ribosomal RNA (rRNA)
• RNA polymerase II
• Transcribes protein-coding genes to make
messenger RNA (mRNA)
• RNA polymerase III
• Transcribes 5S rRNA, transfer RNA (tRNA), and
small nuclear RNA genes (snRNA)
Initiation
• Promoter complex
• TATA box, CAAT box, GC-
rich boxes, octamer
boxes
• Transcription factors
recognize the
promoter and
promote transcription
• RNA polymerase II
then binds and forms
the transcription
initiation complex.
Elongation
• mRNA is synthesized 5’ to 3’
• The FACT complex reassembles the
nucleosomes as polymerase synthesizes the
mRNA
Termination
• RNA polymerase I requires a termination protein
that recognizes a specific 18 nucleotide sequence.
• RNA polymerase III terminates transcription via
hairpin formation.
• RNA polymerase II transcribes 1,000 – 2,000
nucleotides excess of the gene template which are
removed during mRNA processing.
Pre-mRNA Modifications in Eukaryotes
• Precursor-mRNA (pre-mRNA)
-Must be processed in nucleus to produce
translatable mRNA
1. 5´ cap
-guanine nucleotide added to 5’ end
-Protects mRNA from RNA-digesting enzymes
-Site where ribosome attaches to mRNA during
translation
2. Addition of Poly(A) tail
-50 to 250 adenine nucleotides added to 3´ end
-Protects mRNA from RNA-digesting enzymes
3. Removal of Introns
-Non-protein-coding sequences in the pre-mRNA
4. Exons
-Amino acid coding sequences in pre-mRNA
-Joined together sequentially in final mRNA
Pre-mRNA Splicing
• Pre-mRNA splicing involves the
precise removal of introns from
the primary RNA transcript.
• The splicing process is catalyzed
by protein complexes called
spliceosomes that are
composed of proteins and RNA
molecules called snRNAs.
• Spliceosomes recognize
sequences at the 5' and 3' end
of the intron.
Transcription: Initiation
Transcription: Elongation
Transcription: Termination
Take Home Message
q Transcription is the first step in the two-step
process of producing proteins based on
instructions contained in DNA.
q In transcription (which occurs in the nucleus of
eukaryotic cells), a single copy of one specific gene
in the DNA is made in the form of a molecule of
mRNA.
q When the mRNA copy of a gene is completed and
processed, it moves to the cytoplasm, where it can
be translated into a polypeptide.
Transcription Videos
Amobea Sisters video of both transcription and translation.
[Link]
wL0Myd7Dk1F0iQPGrjehze3eDpco1eVz&t=0s
[Link]
resources/dna-transcription-advanced-detail