Polynucleotide Phosphorylase

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?


All figure & text material ©2026 by Steven M. Carr