Archaeal cytoskeletal proteins play pivotal roles in cellular morphology and division, yet their interactions and regulatory networks remain largely unexplored. In this study, we investigated the interactome of archaeal cytoskeletal proteins with known functions involved in cell division and cell shape to explore the interactions between these proteins.
We conducted pull-down experiments using HA and Strep II affinity tags, optimizing elution conditions for enhanced specificity and yield. Western blot validation ensured the specificity of identified interactions. Comparative analysis of tag efficiencies was performed to understand their impact on the interactome study.
Our results revealed a comprehensive network of interactions involving many cytoskeletal proteins. For example, the cell shape control protein CetZ1 exhibited prominent associations with FtsZ2, a cell division protein essential for cytokinesis. Additionally, we discovered interactions with a novel actin-like protein, Volactin, further illuminating the diversity of archaeal cytoskeletal interactors. Our findings also unveiled associations with FtsZ1, another cell division protein, and several uncharacterized proteins, expanding the repertoire of potential CetZ1 interactors. Intriguingly, our study highlighted the involvement of proteins in transcription and translation processes, suggesting a broader cellular regulatory network associated with CetZ1 function.
This research not only delves into the uncharted territory of archaeal cytoskeletal proteins but also sheds light on their intricate interactions. By comparing tag efficiencies, we have extended our understanding of CetZ1 interactors, offering valuable insights into archaeal cell biology. Our findings contribute to the field of archaeal biology and provide a foundation for further investigations, enhancing our comprehension of cellular organization and division in these ancient microorganisms.