THE OVERALL OBJECTIVE OF THIS APPLICATION IS TO DEMONSTRATE THAT SYNTHESIS OF MACROMOLECULES CAN BE ACHIEVED USING ONLY PREBIOTIC MATERIALS: TRIAZINES SH2 AMINO ACIDS FORMALDEHYDE AND SUGARS. THE CENTRAL HYPOTHESIS OF THIS APPLICATION IS THAT TRIAZINES SERVED AS PRECURSORS TO THE NATIVE NUCLEOBASES. OUR RATIONALE FOR THIS HYPOTHESIS IS AS FOLLOWS: 1) TRIAZINES ARE READILY FORMED FROM PREBIOTIC CONDITIONS; 2) TRIAZINES HAVE A HIGH PROPENSITY FOR ASSEMBLY; 3) TRIAZINES ARE FUNCTIONALLY AND STRUCTURALLY COMPATIBLE WITH DNA AND RNA MOLECULES IN EXTANT BIOLOGY. NUCLEOBASE-RECOGNITION IS REQUIRED FOR BIOSYNTHESIS OF SUGARS LIPIDS PROTEINS AND NUCLEIC ACIDS. THE CENTRAL GOAL OF THIS PROPOSAL IS TO EXPLORE THE POSSIBILITY THAT TRIAZINES CAN FULFILL THE NUCLEOBASE ROLE IN RECOGNITION-DRIVEN SYNTHESIS OF MACROMOLECULES. NOTABLY PREBIOTICALLY PLAUSIBLE MACROMOLECULES THAT INTERFACE WITH EXTANT NUCLEIC ACIDS ARE THOUGHT TO BE CENTRAL IN THE ORIGIN OF LIFE. WE WILL TEST OUR HYPOTHESIS OF TRIAZINE FUNCTION AND BY DOING SO ACCOMPLISH OUR CENTRAL GOAL VIA THE FOLLOWING THREE SPECIFIC OBJECTIVES: 1. SYNTHESIS OF ACTIVATED TRIAZINE AND AMINO ACID MONOMERS. THE WORKING HYPOTHESIS OF THIS OBJECTIVE IS THAT THE MATERIALS AVAILABLE IN THE PREBIOTIC WORLD WERE SUFFICIENT FOR THE GENERATION OF MACROMOLECULES THROUGH MONOMER ACTIVATION. NEITHER TRIAZINE NOR AMINO ACIDS AND SUGARS ARE SUFFICIENTLY REACTIVE TO UNDERGO EFFICIENT OLIGOMERIZATION WITHOUT ACTIVATION. THE GOAL OF THIS OBJECTIVE IS TO DEVELOP A PREBIOTIC SYNTHESIS OF THIOESTER-ACTIVATED AMINO ACIDS AND TRIAZINE-GLYCOCONJUGATES. TRIAZINE AND CARBOXYLIC ACID ACTIVATION WILL BE PURSUED USING PREBIOTICALLY PLAUSIBLE SULFUR MINERAL AND FORMALDEHYDE CHEMISTRY. 2. TEMPLATED OLIGOMERIZATION OF TRIAZINE AND AMINO ACIDS. THE WORKING HYPOTHESIS OF THIS OBJECTIVE IS THAT THE SELF AND PAIR-WISE ASSEMBLING PROPERTIES OF TRIAZINES WOULD ENABLE THE OLIGOMERIZATION OF ACTIVATED MONOMERS EVEN AT LOW OVERALL CONCENTRATION. THE GOAL OF THIS OBJECTIVE IS THE QUANTITATIVE EVALUATION OF TRIAZINE ASSEMBLY-DIRECTED OLIGOMER SYNTHESIS. IN PARTICULAR OLIGOMERIZATION OF A BINARY MIXTURE OF HYDROPHOBIC AND POLAR AMINO ACIDS WILL BE STUDIED TO EXAMINE THE INFLUENCE AND EMERGENCE OF AMPHIPATHIC CHARACTER DURING THE SYNTHESIS OF PEPTIDE OLIGOMERS IN WATER. 3. DNA AND RNA TEMPLATED SYNTHESIS OF PEPTIDE AND TRIAZINE MACROMOLECULES. THE WORKING HYPOTHESIS OF THIS OBJECTIVE IS THAT THE TRIAZINE ASSEMBLY-TEMPLATED CHEMISTRY OF PREBIOTICALLY-ACTIVATED MONOMERS WILL BE COMPATIBLE WITH EXTANT BIOMACROMOLECULES SUCH AS DNA AND RNA. THE GOAL OF THIS OBJECTIVE IS TO FIND CONDITIONS FOR DNA AND RNA TEMPLATED OLIGOMERIZATION OF PREBIOTICALLY ACTIVATED MONOMERS. THE SUCCESSFUL COMPLETION OF THIS GOAL WOULD SUPPORT THE NOTION THAT TRIAZINES COULD SERVE AS A CHEMICAL INTERMEDIARY BETWEEN THE PREBIOTIC AND RNA WORLDS. SIGNIFICANCE THIS PROPOSAL DIRECTLY ADDRESSES PROGRAM ELEMENT SUBHEADING PREBIOTIC EVOLUTION: DETERMINING WHAT CHEMICAL SYSTEMS COULD HAVE SERVED AS PRECURSORS OF METABOLIC AND REPLICATING SYSTEMS ON EARTH AND ELSEWHERE INCLUDING ALTERNATIVES TO THE CURRENT DNA-RNAPROTEIN BASIS FOR LIFE. THE OBJECTIVES ARE FRAMED BY A HYPOTHESIS THAT TRIAZINES WERE FUNCTIONAL PRECURSORS TO THE NATIVE BASES; EXTENSION OF OUR HYPOTHESIS SUGGESTS THAT TRIAZINES COULD LIKEWISE DIRECT COUPLING OF BIOLOGICALLY IMPORTANT MONOMERS. THIS PROPOSAL WILL FOCUS ON THE USE OF TRIAZINE ASSEMBLY-MOTIFS TO DRIVE THE FORMATION OF AMINO ACID AND SUGAR-DERIVED POLYMERS UNDER PREBIOTICALLY RELEVANT ENZYME-FREE CONDITIONS. THERE ARE NO ALTERNATIVE PROGRAM ELEMENTS BETTER SUITED TO THIS PROPOSAL.