From Gene to Protein

Transcription and Translation

2007-2008

What do genes code for?

How does DNA code for cells & bodies?

how are cells and bodies made from the instructions in DNA

DNA

proteins

cells

bodies

The Central Dogma

Flow of genetic information in a cell
How do we move information from DNA to proteins?

DNA

RNA

protein

trait

replication

2007-2008

RNA
ribose sugar N-bases
uracil instead of thymine U:A C:G

single stranded lots of RNAs

mRNA, tRNA, rRNA, siRNA

DNA

transcription

RNA

Transcription
Making mRNA
enzyme
RNA polymerase

5
C

DNA

A G T A T C T A

G C

A G C T

C G T

T A

C A G C T G

A T

A T

3 5

3
G C A U C G U C G T A G C A T

unwinding

rewinding

mRNA

build RNA 53

RNA polymerase

RNA polymerases
3 RNA polymerase enzymes
RNA polymerase 1
only transcribes rRNA genes makes ribosomes

RNA polymerase 2
transcribes genes into mRNA

RNA polymerase 3
only transcribes tRNA genes

Matching bases of DNA & RNA

Match RNA bases to DNA bases on one of the DNA strands
AG C A G U G U C G G A U A

C A

U C

C
A U

5'

RNA A C C polymerase G

3'

T GG T A C A G C T A G T C A T CG T A C CG T

Eukaryotic genes have nucleotide sequences that do not code for anything

Eukaryotic genes are not continuous

exons = the real gene
expressed / coding DNA

introns = the junk

inbetween sequence

intron = noncoding (inbetween) sequence

eukaryotic DNA
exon = coding (expressed) sequence

mRNA splicing
Post-transcriptional processing
eukaryotic mRNA needs work after transcription primary transcript = pre-mRNA mRNA splicing edit out introns make mature mRNA transcript
intron = noncoding (inbetween) sequence

~10,000 base

eukaryotic DNA
exon = coding (expressed) sequence

pre-mRNA

primary mRNA transcript mature mRNA transcript spliced mRNA

~1,000 base

RNA splicing enzymes

snRNPs
small nuclear RNA exon proteins
snRNA intron snRNPs exon 3'

Spliceosome

5'

several snRNPs recognize splice site sequence

cut & paste gene

spliceosome 5' 3' lariat 5' 3'

mature mRNA

exon 5'

exon 3' excised intron

Alternative Splicing
results in a single gene coding for multiple proteins

 Need to protect mRNA on its trip from nucleus to cytoplasm

enzymes in cytoplasm attack mRNA
protect the ends of the molecule add 5 GTP cap add poly-A tail
longer tail, mRNA lasts longer: produces more protein

3' A

mRNA P G P P

5'

mRNA codes for proteins in triplets

DNA

TACGCACATTTACGTACGCGG
codon

mRNA AUGCGUGUAAAUGCAUGCGCC

?
protein

Met Arg Val Asn Ala Cys Ala

 Code for ALL life!

strongest support for a common origin for all life

Code is redundant
several codons for each amino acid 3rd base wobble

Start codon

AUG methionine

Stop codons

UGA, UAA, UAG

How are the codons matched to amino acids?

DNA

TACGCACATTTACGTACGCGG
5 3

mRNA AUGCGUGUAAAUGCAUGCGCC
3 5

codon

UAC
tRNA amino acid

GCA CAU anti-codon Met

Arg

Val

Transfer RNA structure

Clover leaf structure
anticodon on clover leaf end amino acid attached on 3 end

Ribosomes
Facilitate coupling of tRNA anticodon to mRNA codon Structure
ribosomal RNA (rRNA) & proteins 2 subunits
large small
E P A

Building a polypeptide
Initiation
brings together mRNA, ribosome subunits, initiator tRNA

Elongation
adding amino acids based on codon sequence

Termination
end codon
Leu Met Met tRNA Met Leu Met

3 2 1
Val
Leu
Leu

Ser

release factor Trp

Ala

U AC 5' C UGAA U mRNA A U G 3' E P A

5'

UAC GAC A U G C U GA A U

5'
3'

U A C GA C AA U A U G C UG

5' 3'

U AC G A C AA U AU G C UG

3'

A CC U GG U A A

3'

Translation simulation John Kyrk

Protein Targeting
Signal peptide
address label
start of a secretory pathway

Destinations:

secretion nucleus mitochondria chloroplasts cell membrane cytoplasm etc

RNA polymerase DNA

REVIEW
exon intron

amino acids tRNA

5' GTP cap

pre-mRNA mature mRNA

poly-A tail large ribosomal subunit

aminoacyl tRNA synthetase

polypeptide

3'

5'
small ribosomal subunit

tRNA E P A

ribosome

The Transcriptional unit (gene?)

enhancer
1000+b

20-30b 3' RNA TATA polymerase DNA

translation start TAC

exons

translation stop ACT 5' DNA

transcriptional unit (gene)

introns promoter transcription start

5' pre-mRNA

transcription stop
3'

5' GTP mature mRNA

3'

AAAAAAAA

Bacterial chromosome

Protein Synthesis in Prokaryotes

Cell membrane Cell wall
2007-2008

Transcription mRNA

Prokaryote vs. Eukaryote genes

Prokaryotes
DNA in cytoplasm circular chromosome naked DNA no introns transcription and translation at same time

Eukaryotes
DNA in nucleus linear chromosomes DNA wound on histone proteins introns vs. exons transcription and translation separate in time

Can you tell the story?

The Transcriptional unit

enhancer
1000+b

20-30b 3' RNA TATA polymerase TAC

exons transcriptional unit

ACT 5' DNA

introns

5'

3'

5'

3'