Outline One enzyme ?? One gene Structure and function of proteins

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Sections & The genetic code is _________, meaning that an amino acid may be coded by more than one codon. unambiguous degenerate.

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Outline One enzyme ?? One gene Structure and function of proteins Genetic Code Process of translation Iniation Elongation Termination Release modification One enzyme ??

Prototrophs mutation Auxotrophs

The Structure and Function of Proteins

Amino Acids-R groups Nonpolar, aliphatic Polar, uncharged Aromatic Positively charged Negatively charged

Protein: Levels of structural organization

The Genetic Code and Translation

The Breaking of the Genetic Code

Experiments that “broke the genetic code” 1. Homopolymer experiment 2. Known RNA sequences and ribosomal bound tRNA

1. . 3 2 1 All these experiments used a cell free system. 4 5 ppt. protein

1. Homopolymer experiment In 1961 1 hot amino acid translation 19 cold amino acids Cell free system 20X each a different “hot amino acid

2. Known mRNA sequences + Ribosomal bound tRNA Amino acid This is transfer RNA Anti-codon

Amino Acid Transfer RNA ribosome

2. Known mRNA sequences + Ribosomal bound tRNA Amino acid This is transfer RNA Anti-codon

Isolate ribosome bound tRNA Mix with Codon of mRNA Isolate ribosome bound tRNA Analyze amino acid

Genetic Code Table 1968 Wobble Degeneracy & mRNA

Isoaccepting & tRNAs Different tRNAs accept same amino acid!

The process of translation

mRNA sequences

Translation 1. Binding of amino acids to tRNA 2. Initiation 3. Elongation 4. Termination 5. Peptide Release 6. Protein modification

Step 1 Binding of amino acid to tRNA

It takes two steps to “charge” the tRNA Aminoacyl-tRNA-synthetase Amino acid+ATP tRNA is charged

Step 2: Initiation of Translation Bacterial Initiation IF3

30S

Note 3 ribosomal sites E P A 70S E A

Eukaryotic Initiation Kozak Eukaryotic Initiation key differences

3. ELONGATION: look at the “EPA” sites

Elongation continues on ribosomes

Steps 4 and 5: UAA UAG UGA Termination and Release

6. Lipoprotein Glycoprotein

Many antibiotics target prokaryotic translation.

Antibiotics and Translation Bind and block bacterial ribosomes “A” site Binds to large subunit; blocks peptide formation Blocks initiation step Blocks translocation step Tetracyclines Chloramphenicol Streptoymycin Erythromycin

Summary of important steps involved in translation For your review: Summary of important steps involved in translation

1. 2.

3.

4.

5.

6. Protein Modifications

Eukaryotic translation is very similar except: More IF’s Kozak sequence 5’CAP 3’poly-A tail