Transcription: a primer

(eukaryotic)

Aurnab Ghose; 150202

Same genome!

Same genome!

Anise swallowtail, Papilio zelicaon

Gene

Translation at ribosome

mRNA

Transcription

Protein

Transcription

Requires

Transcription initiation site

where transcription begins

Promoter

to which RNA polymerase binds

In multicellular eukaryotes

RNA polymerase binds to promoter (TATA
box)

TATA box
UPE

Transcription
initiation
site

T T
TATA A
A A

pre-mRNA

Eukaryotic promoter elements. A eukaryotic

promoter usually contains a TATA box located 25-35
bp upstream from the transcription initiation site. One
or more UPEs (upstream promoter elements) is
usually present.

Enhancers

Located far away from promoter

control some gene expression

Help form active transcription initiation

complex

Specific regulatory proteins

bind to enhancer elements

activate transcription by interacting with
proteins bound to promoters

Enhancers

actcaaaaaaaaaacggttgggttgcgccatacatatgaaag
agtatagaataatgatgtatttcccaaatcaaatatcatggt
aaaatttaaCAATgacccattcggattcattgataatatta
gttgatggatcatttgtaaaaaggttttattaactcctaagt
tatgtcgagtagaccttgttgttgttgcTATAATTcttaa
tcATGcgttgtagggggagatttatgtcaccacaaacagaa
acgaaagcaaaggttgggttcaaagctggtgttaaagactga
acgtagcagctacgatcgatcgactagctgcatcgggctagc
gaagcttcgatcgatcgatcgagctagcgagcccccagtttt
aggtcgagctttcagctcagctaggcgcgaaatctcgagcgc
agctcactagctgctctagcatcgagctacgatcgcgatcga
gctagctagaattatccgtgaagcttgcaaatggagtcctga
attagctgctgcttgtgaagtctggaaggaaatcaaatttga
attcccagcaatggatactttgTAAtccagtaataatcat
tcgttctattaatttccattaaactcggcccaatctt

Transcription

Is the process of making messenger RNA
(mRNA) from a DNA template
RNA polymerase
Very similar to DNA replica;on
Remember: as in replica;on, in transcrip;on,
addi0on of a new nucleo0de occurs at the 3
end!
Transcrip;on occurs by base pairing:
A-U: 2 H bonds; G-C: 3 H bonds

Three Steps of transcription

Transcrip;on
Complementary Base-pairing of DNA to mRNA

If the template strand reads:

T-A-C-C-T-T-A-A-C-C-G-G-T-T-A
The transcribed mRNA is:
A-U-G-G-A-A-U-U-G-G-C-C-A-A-U

The structure of mRNA

1. 5 Guanine cap (G-cap)

2. Leader sequence: does not get made into protein
3. Protein coding region begins with AUG & ends with a STOP
codon
4. Trailing sequence does not get made into protein.
5. 3 poly-A tail

1. 5 Guanine cap (G-cap)

2. Leader sequence: does not get made into protein

3. Protein coding region begins with AUG & ends with
a STOP codon

4. Trailing sequence does not get made into protein.

5. 3 poly-A tail

Gene Expression: Translation

Central Dogma: DNA RNA Protein

Proposed by Francis Crick
DNA:
3' ACC AAA CCG AGT
mRNA: 5' UGG UUU GGC UCA
Protein: Trp Phe Gly Ser

The string of amino acids has a direct rela;onship to

nucleo;de bases in RNA and DNA
Every three nucleo;des is called a Codon
Each Codon corresponds to ONE amino acid

 Amino acid subunits are added at the carboxyl

terminus of the growing protein

The elements of eukaryotic

transcription

Cis-acting elements

Basal promoter elements

UPEs

Enhancers

Trans-acting factors

RNA polymerase

Transcription factors

Accessory proteins

Modulating eukaryotic
transcription

Promoter proximal elements (UPEs)
Enhancer elements: Can function in either orientation.
Can occur far (<50 kb) from the gene.
Can be up- or downstream.

Three RNA polymerases in

eukaryotes"

I: pre-rRNA"
II: mRNA"
III: tRNAs, 5S rRNA, "
small stable RNAs"
"
!
Also differentiated by
sensitivity to -amanitin.!

Complexity of eukaryotic RNA

polymerases"

Function of most
subunits is unclear, but
nearly all the subunits
are essential

Transcription initiation at eukaryotic

protein-coding genes

# of subunits
RNA pol II

Activity

12

RNA synthesis

TFIID

Promoter recognition

TFIIB

TFIIE

TFIIF

TFIIH

Mediator

20

General TX factors

helicase

TBP binds to TATA-box DNA"

TATA-binding protein = TBP!

!
TBP is a subunit of TFIID!

Pol II won t bind unless TFs bind first.

Stepwise assembly of Pol II

transcription-initiation complex"

Transcriptional control requires

still more proteins

RNA pol II (12 proteins)
General transcription factors (23 proteins)
chromatin remodeling complex (15 proteins)
specific transcription factors (? proteins)
Mediator complex (20 proteins)