Protein synthesis using poly(2'-halogeno-2'-deoxyadenylic acids) as messenger

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Nucleic Acids

Background:

• Messenger RNAs (mRNAs) are crucial for protein synthesis, directing the sequence of amino acids.
• Synthetic polynucleotides offer a versatile tool for studying the mechanisms of translation.
• Modifications to the sugar moiety of nucleotides can influence their properties and biological activity.

Purpose of the Study:

• To investigate the efficacy of novel synthetic polynucleotides as messenger RNAs in an in vitro protein synthesis system.
• To compare the messenger activity and stability of poly(2"-fluoro-2"-deoxyadenylic acid), poly(2"-chloro-2"-deoxyadenylic acid), and poly(2"-bromo-2"-deoxyadenylic acid) against a standard poly(riboadenylic acid) (poly(rA)).

Main Methods:

• In vitro protein synthesis assays were conducted using cell-free systems.
• Synthetic polynucleotides were employed as messenger RNAs to direct polypeptide synthesis.
• The incorporation of radiolabeled lysine ([14C]lysine) into polypeptides was measured to assess messenger activity.
• Initial velocities of polypeptide formation and the longevity of messenger activity were determined.

Main Results:

• All tested synthetic polynucleotides, including poly(2"-fluoro-2"-deoxyadenylic acid), poly(2"-chloro-2"-deoxyadenylic acid), and poly(2"-bromo-2"-deoxyadenylic acid), were active as messengers in the protein synthesis system.
• [14C]lysine was successfully incorporated into polypeptides directed by these synthetic messengers.
• Poly(2"-fluoro-2"-deoxyadenylic acid) exhibited a greater initial velocity for polylysine formation compared to poly(rA).
• All synthetic messengers demonstrated increased stability and longevity within the protein synthetic system.

Conclusions:

• The modified polynucleotides, particularly poly(2"-fluoro-2"-deoxyadenylic acid), function effectively as synthetic messengers in in vitro translation.
• These novel messengers show improved performance, including higher initial synthesis rates and greater stability, compared to conventional poly(rA).
• The findings highlight the potential of chemically modified nucleic acids in advancing studies of protein synthesis and genetic code.