bioRxiv Science⌕ Search

Biology subjects

Patki, G. M.

Publications and source records attributed to Patki, G. M..

2 recordsLinked to original sources

Characterizing mixed single chain amphiphile-based coacervates as a robust protocell system

Prebiotic soup would have been a dilute pool of various constituent chemicals that would have reacted with each other to form biologically relevant precursors during lifes origin. In this milieu, compartments formed by liquid-liquid phase separation (LLPS) are thought to have facilitated concentration of chemicals, thereby catalyzing their reactions. Towards this, various LLPS-based systems have been studied as model protocells. Relevantly, fatty acid-based (decanoic acid) coacervates have recently been explored as model protocells. As far as protocell research is concerned, fatty acids have been studied much more extensively in the context of vesicle-forming entities when compared to them resulting in coacervate systems. Furthermore, exogenous delivery and endogenous synthesis of fatty acids suggest the prevalence of single chain amphiphiles (SCAs) on the early Earth, with a greater abundance of the shorter chain length moieties. In this backdrop, we set out to fabricate robust coacervate-based protocells using SCAs that would have been readily present in a chemically heterogeneous prebiotic soup, and which could thrive under various prebiotically relevant selection pressures. Towards this, we characterized a mixed amphiphile-based coacervate system composed of nonanoic acid (NA), nonanol (NOH) and tyramine (Tyra), which could form coacervates over a broad range of pHs, temperatures, and salt concentrations. This is noteworthy as compositionally heterogenous vesicles have also been shown to have advantages over pure fatty acid vesicles. Additionally, we also demonstrate RNA sequestration in these coacervates that gets enhanced upon addition of cationic amino acids, emphasizing the importance of co-solute interactions in the prebiotic soup. Lastly, we also demonstrate nonenzymatic template-directed primer extension in these coacervates, suggesting the potential functional role of these compartments during lifes origin.

biochemistry↗

Effect of 'spent'nucleotides' on nonenzymatic RNA replication

Nonenzymatic template-directed replication would have been affected by co-solutes in a heterogenous prebiotic soup due to lack of enzymatic machinery. Unlike in contemporary biology, these reactions use chemically-activated nucleotides, which undergo rapid hydrolysis forming nucleoside monophosphates ( spent monomers). These co-solutes cannot extend the primer but continue to base pair with the template, thereby interfering with replication. We therefore aimed to understand how a mixture of spent ribonucleotides would affect nonenzymatic replication. We observed inhibition of replication in presence of the mixture, wherein predominant contribution came from the cognate Watson-Crick monomer, showing potential sequence dependence. Our study highlights how nonenzymatic RNA replication would have been directly affected by co-solutes, with ramifications for the emergence of functional polymers in an RNA World.

biochemistry↗