Ch 15 (this will be also discussed in lab)
Transcript of Ch 15 (this will be also discussed in lab)
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Genetic Code and Translation
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Outline
The Breaking of the Genetic Code Examples of two experimental approaches
Concepts and Terms related to genetic code Wobble Degeneracy isoaccepting
Translation prokaryote eukaryotes
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The Genetic Code and Translation
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The Breaking of the Genetic Code
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Experiments that “broke the genetic code”
1. Cell Free experiments Applied to the Homopolymer experiment
2. Known RNA sequences and ribosomal bound tRNA
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All these experiments used a cell free system.
1 23
4
ppt. protein5
1. Overview
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Cell free system
1 hot amino acid
19 cold amino acids
translation
20X each a different “hot amino acid
Application of experimental design
+
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2. Known mRNA sequences + Ribosomal bound tRNA
This is transfer RNA
Anti-codon
Amino acid
Now link this idea toThe genetic code table
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Transfer RNA
Amino Acid
ribosome
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Codon of mRNAMix with
Isolate ribosome bound tRNA Analyze amino acid
2. EXPERIMENT
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Close up of a possible result:
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Genetic Code Table1968
Degeneracy & mRNA
Wobble
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Isoaccepting & tRNAs
Different tRNAs accept same amino acid!
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The process of translation
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mRNA sequences
We will approach our discussion of translation as a “Nonoverlapping Code”
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Steps of Translation
1. Binding of amino acids to tRNA
2. Initiation
3. Elongation
4. Termination
5. Peptide Release
6. Protein modification
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Step 1 Binding of amino acid to tRNA
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It takes two steps to “charge” the tRNA
Amino acid+ATPtRNA is charged
Aminoacyl-tRNA-synthetase
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Up close view of “charged” tRNA
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Step 2: Initiation of Translation
IF3
Bacterial Initiation
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30S
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70S
Note 3 ribosomal sites E P A
E A
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Kozak
Eukaryotic Initiationkey differences
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3. ELONGATION: look at the “EPA” sites
Peptidyl transferase
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Up close view of translation
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Elongation continues on ribosomes
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Termination and Release
UAAUAGUGA
Steps 4 and 5:
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Animation of Translation
http://highered.mcgraw-hill.com/sites/0072437316/student_view0/chapter14/animations.html
See chapter 15
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6.
Lipoprotein
Glycoprotein
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Many antibiotics target prokaryotic translation.
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Antibiotics and Translation
Tetracyclines
Chloramphenicol
Streptoymycin
Erythromycin
Bind and block bacterial ribosomes “A” site
Binds to large subunit; blocks peptide formation
Blocks initiation step
Blocks translocation step
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Review: The Structure and Function of Proteins
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Amino Acids-R groups
Nonpolar, aliphatic
Polar, uncharged
Aromatic
Positively charged
Negatively charged
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Protein: Levels of structural organization
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For your review:Summary of important steps involved in translation
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1.
2.
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3.
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4.
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5.
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6. Protein Modifications
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Eukaryotic translation is very similar except:
More IF’s
Kozak sequence
5’CAP
3’poly-A tail
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