Cracking the Genetic Code 

The Central Dogma: DNA makes RNA makes protein
Central Dogma

After 1953: DNA somehow "codes" for proteins
    Proteins are made on ribosomes , which contain
         long-lived (stable) & short-lived (labile) RNA  

Deciphering the code
       Can't be 1- or 2-letter code:
            1 letter 4 amino acids only; 2 letters  4 x 4  =  16 only
           20 amino acids require a minimum 3-letter triplet code
                 4 x 4 x 4 = 64, is there redundancy? (hmmm ....)

        Can't overlap: neighbors aren't constrained
                                 point mutations affect only single amino acids

        Can't be direct DNA protein:
            not chemically compatible (H-bonds vs. covalent bonds)
            in eukaryotes, DNA & protein are physically separated

       Adaptor Hypothesis (Crick 1957)
                some molecule must serve as intermediary

       Messenger RNA  hypothesis (Jacob & Monod 1961)
           rRNA is too stable; "other RNA" is labile (transient)
                "other RNA" acts as messenger: mRNA is the "blueprint"
                rRNA is the "workshop"

Experiments with RNA homopolymers (Nirenberg 1961)

    cell-free in vitro protein synthesis:
       polynucleotide phosphorylase polymerizes rNDP monomers into RNA
                 rNDP  +  rNDP  + ... +  rNDP  <=>  3'-rNM-rNM-...-rNMP-5' + nPi

    UUU UUU UUU        poly-phenylalanine, therefore    UUU codes phe
    AAA AAA AAA       
poly-lysine                                     AAA codes lys
    CCC CCC CCC       
poly-proline                                   CCC codes pro
    GGG GGG GGG     
poly-glycine                                  GGG codes gly

Experiments with RNA di-, tri-, & tetra-nucleotide polymers (Khorana 1965)

(1) UG + UG + UG + UG   UGU-GUG-UGU-GUG 
            cysteine +
valine + cysteine + valine + . . . . etc

                                                       GGU-GGU-GGU or

        mixture of poly-trypotophan, poly-glycine, & poly-valine

      GUG & valine occur & produced in both experiments
            therefore, GUG makes valine

                        trp, val, gly in 1:1:2 ratio

        Since we know GUG makes valine,
           and we know GGG makes glycine,
                               if GGU makes gly as well (hmmm...),
        Therefore, UGG makes trypotophan
                & maybe GGN makes glycine: code is "ambiguous" in 3rd position

     and so on for all 64 triplets

The complete code was deciphered by 1965 (Nobel prize 1968)
    The code is a logical relationship between DNA, RNA, & protein
    The code precedes the biochemical understanding of transcription & translation

    [First example of Mega-Project Biology: NIH organized major effort through Nirenberg] 

All text material ©2014 by Steven M. Carr 

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