1. GENE EXPRESSION
© 2016 Paul Billiet ODWS

2. Two steps are required
Transcription The synthesis of mRNA uses the gene on the DNA molecule as a template This happens in the nucleus of eukaryotes
Translation The synthesis of a polypeptide chain using the genetic code on the mRNA molecule as its guide.
© 2016 Paul Billiet ODWS

3. RIBONUCLEIC ACID (RNA)
Found all over the cell (nucleus, mitochondria, chloroplasts, ribosomes and the soluble part of the cytoplasm).
© 2016 Paul Billiet ODWS

4. Types Messenger RNA (mRNA) <5% Ribosomal RNA (rRNA) Up to 80%
Transfer RNA (tRNA) About 15%
In eukaryotes small nuclear ribonucleoproteins (snRNP aka spliceosomes)
© 2016 Paul Billiet ODWS

5. Structural characteristics of RNA molecules
Single polynucleotide strand which may be looped or coiled (not a double helix)
Sugar Ribose (not deoxyribose)
Bases used: Adenine, Guanine, Cytosine and Uracil (not Thymine).
© 2016 Paul Billiet ODWS

6. mRNA A long molecule 1 million Daltons Ephemeral Difficult to isolate
mRNA provides the plan for the polypeptide chain
© 2016 Paul Billiet ODWS

7. rRNA
Coiled
Two subunits: a long molecule 1 million Daltons a short molecule Daltons
Fairly stable
Found in ribosomes
Made as subunits in the nucleolus
rRNA provides the platform for protein synthesis
© 2016 Paul Billiet ODWS

8. tRNA Short molecule about 25 000 Daltons Soluble
At least 61 different forms. Each, have a specific anticodon as part of their structure.
tRNA “translates” the message on the mRNA into a polypeptide chain
© 2016 Paul Billiet ODWS

9. snRNA Spliceosomes Small sequences of RNA that can cut mRNA
They edit mRNA in eukaryotes
Remove non-coding parts (introns)
© 2016 Paul Billiet ODWS

10. Transcription: The synthesis of a strand of mRNA (and other RNAs)
Uses an enzyme RNA polymerase
Proceeds in the same direction as replication (5’ to 3’)
Forms a complementary strand of mRNA
It begins at a promotor site, which signals that the beginning of the gene is near (about 20 to 30 nucleotides away)
After the end of the gene is reached, there is a terminator sequence that tells RNA polymerase to stop transcribing
NB Terminator sequence ≠ terminator codon.
© 2016 Paul Billiet ODWS

11. RNA polymerase
© 2016 Paul Billiet ODWS

12. Editing the mRNA
In prokaryotes, transcribed mRNA goes straight to the ribosomes in the cytoplasm
In eukaryotes, freshly transcribed mRNA in the nucleus is about 5000 nucleotides long
When the same mRNA is used for translation at the ribosome it is only 1000 nucleotides long
The mRNA has been edited
The parts which are kept for gene expression are called EXONS (exons = expressed)
The parts which are edited out (by spliceosomes) are called INTRONS.
© 2016 Paul Billiet ODWS

13. Spliceosome from yeast
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14. Transcription plan Nucleus Gene DNA Transcription messenger RNA
© 2016 Paul Billiet ODWS

15. Translation plan Complete protein Polypeptide chain TRANSLATION
Ribosomes
Stop codon
Start codon
© 2016 Paul Billiet ODWS

16. Translation
Location: The ribosomes in the cytoplasm that provide the environment for translation
The genetic code is brought by the mRNA molecule.
© 2016 Paul Billiet ODWS

17. What is the genetic code?
The genetic code = the sequence of bases found along the mRNA molecule
There are only four letters to this code (A, G, C and U)
The code needs to be complex enough to represent 20 different amino acids used to build proteins.
© 2016 Paul Billiet ODWS

18. How many combinations?
If one base represented one amino acid this would only be able to produce…
4 different combinations. (A, C, G and U)
If pairs of bases represented each amino acid this would only be able to produce…
4 x 4 = 16 combinations. (AA, AC, AG, AU, CA, CC, CG, CU etc)
If triplets of bases represented each amino acid, this would be able to produce…
4 x 4 x 4 = 64 combinations This is enough combinations to code for the 20 amino acids but is the code actually made of triplets?
© 2016 Paul Billiet ODWS

19. Nature is logical!
Over 10 years biochemists synthesised bits of mRNA with different combinations
Then they used them to synthesise polypeptides
The results proved the logical answer was correct
The genetic code is made of triplets of bases called codons.
© 2016 Paul Billiet ODWS

20. The Central Dogma Proposed by Francis Crick 1958
DNA holds the coded hereditary information in the nucleus
This code is expressed at the ribosome during protein synthesis in the cytoplasm
The protein produced by the genetic information which is influenced by natural selection
If a protein is modified it cannot influence the gene that codes for it
Therefore there is a one way flow of information:
DNARNAProtein
© 2016 Paul Billiet ODWS

21. An important discovery
Retro viruses (e.g. HIV) carry RNA as their genetic information
When they invade their host cell they convert their RNA into a DNA copy using reverse transcriptase
Reverse transcriptase
Thus the central dogma is modified:
DNA↔RNAProtein
This has helped to explain an important paradox in the evolution of life.
© 2016 Paul Billiet ODWS

22. The paradox of DNA DNA is a very stable molecule
It is a good medium for storing genetic material but…
DNA can do nothing for itself
It requires enzymes for replication
It requires enzymes for gene expression
The information in DNA is required to synthesise enzymes (proteins) but enzymes are require to make DNA function
Which came first in the origin of life DNA or enzymes?
© 2016 Paul Billiet ODWS

23. RIBOZYMES: Both genetic and catalytic
Certain forms of RNA have catalytic properties
RIBOZYMES
Ribosomes and spliceosomes are ribozymes
RNA could have been the first genetic information synthesizing proteins…
…and at the same time a biocatalyst
Reverse transcriptase provides the possibility of producing DNA copies from RNA.
© 2016 Paul Billiet ODWS

24. The ribosome a ribozyme