Synthesis
of Poly-ribonucleotide templates
with Polynucleotide Phosphorylase
Nirenberg &
Matthei (1961) synthesized artificial poly-ribonucleotides
(RNAs) from a pool of monomers of each of the four
available ribonucleoside diphosphates (rADP,
rCDP, rGDP, & rUDP). The enzyme Polynucleotide
Phosphorylase unites the monomers into a long-chain RNA
molecule by removing the terminal phosphate group from each of
the rNDPs, so that the 5'-monophosphate end
can join to the 3' end of the ribose sugar in the
next rNDP. (In the left-hand diagram, the 5'3'
synthesis proceeds right to left). This creates a polynucleotide
RNA (rNMP]n) molecule consisting
entirely of only one species of rNDP. This serves in
effect as a "messenger" RNA in a cell-free
protein synthesis experiment. This first experiment demonstrated
that a 5' 3'poly-UUUUUU template
directed the synthesis of a poly-phenylalanine (F)
amino acid chain, and thus that UUU was the 'genetic
code' for Phe.
HOMEWORK:
The last sentence is not quite accurate: did this first experiment
actually demonstrate a triplet code? Why or why
not?