A myriad of cellular processes such as cell division, migration, wound healing, and muscle cells contraction are dependent on biological nanomachines. One such machinery, called the actomyosin ring network, comprising actin filaments (F-actin), the molecular motor Myosin-II, Tropomyosin, and actin cross-linkers, through their intricate interplay orchestrate cell division. Though the actomyosin machinery is conserved across all eukaryotes – from yeast to humans – a comprehensive understanding of the underlying mechanisms that drive this machinery is still sought after. However, the lack of essential tools necessary to comprehend the dynamics of the process at a cellular and molecular level is a major bottleneck in functionally characterizing the actomyosin ring machinery in multiple biological processes. Thus, to circumvent it, we developed a new nanobody-based toolkit named ALIBY (ALFA nanobody based Toolkit for Imaging and Biochemistry in Yeast). Unlike previously known epitoge tags, the ALFA-tag/NbALFA system is unique in terms of offering a complete set of functionalities including advanced microscopy and biochemistry. Now, the use of this system has been extended to visualize polymeric proteins like actin and tropomyosin, and enabled us to visualize actin dynamics for the first time in live-cells. Thus, with this newfound strategy, will elucidate the spatio-temporal dynamics of essential cytoskeletal proteins during cytokinesis at the cell division site.
Saravanan Palani is an Assistant Professor at the Department of Biochemistry, Indian Institute of Science, Bengaluru. He is currently a DBT-Wellcome India Alliance Intermediate Fellow at IISc. He completed his PhD with Prof Gislene Pereira at German cancer research center (DKFZ), Heidelberg, Germany. He did his Post-Doc with Prof Mohan Balasubramanian at University of Warwick. His primary research interest is in the field of cell division and synthetic biology.