Unravelling the role of the A domain and N-terminal alpha-helices of FtsY in archaeal signal recognition particle

Signal recognition particle (SRP) system is critical for protein translocation across membranes in all domains of life. In archaea, this pathway relies on two GTPase proteins, SRP54 and FtsY, which interact with SRP RNA to facilitate the targeting of nascent proteins to the membrane. Although the SRP components in eukaryotes and bacteria are well characterized, the mechanisms underlying SRP-dependent membrane targeting in archaea remain poorly understood, particularly concerning the role of the FtsY N-terminal domains. This study provides an in-depth exploration of the archaeal SRP system, focusing on the N-terminal domains of the FtsY protein and their role in the formation and functionality of the targeting complex (TC). We characterized the minimal structural domains of FtsY required for SRP54 binding and membrane association, demonstrating the critical involvement of the A domain and N-terminal alpha helices in facilitating these processes. The deletion of these domains led to a progressive reduction in the affinity between SRP54 and FtsY, disrupting TC formation and compromising its catalytic efficiency. Molecular dynamics simulations and thermodynamic analyses corroborated these experimental findings, revealing that the A domain is integral to stabilizing TC and enhancing reciprocal GTP hydrolysis. Furthermore, the study showed that membrane association, mediated by the orientation of the A domain and the alpha N1 helix, is essential for stabilizing the interaction between SRP and the membrane. These results shed light on the molecular basis of SRP assembly and membrane targeting in archaea, marking an important advancement in our understanding of the archaeal SRP machinery.