ABSTRACT
Background: The eukaryotic organism Komagataella pastoris has emerged as a valuable expression system for the production of recombinant proteins. Despite its significance in biotechnological applications, the understanding of the chromatin organization and histone dynamics in K. pastoris remains limited. Nucleosomes, fundamental units of chromatin, play a crucial role in regulating gene expression, and investigating their dynamics is essential for comprehending the molecular processes underlying cellular function.
Aim: This study aims to provide a comprehensive analysis of nucleosome dynamics in K. pastoris by integrating structural and biochemical approaches. Specifically, our goal is to unravel the functional roles of K. pastoris histones, shedding light on their involvement in chromatin organization and gene regulation.
Methods: We employed a multi-faceted approach, combining X-ray crystallography, cryo-electron microscopy, and biochemical assays to investigate the structural and functional aspects of nucleosomes in K. pastoris. Histones were isolated and characterized, and their interactions with DNA were studied using biophysical techniques. Additionally, we explored the post-translational modifications of histones to understand their impact on nucleosome dynamics.
Results: Our integrated analyses reveal the three-dimensional structures of K. pastoris nucleosomes and provide insights into their dynamic behavior. We identify unique features in the histone composition and modifications that contribute to the stability and functional diversity of nucleosomes in this organism. Furthermore, we elucidate the implications of these findings on the regulation of gene expression in K. pastoris.
Conclusion: This study presents a comprehensive examination of nucleosome dynamics in Komagataella pastoris, uncovering crucial insights into the functional roles of histones in chromatin organization. The identified structural features and post-translational modifications contribute to a deeper understanding of gene regulation mechanisms in this biotechnologically important organism. These findings have implications for optimizing protein expression in K. pastoris and offer valuable knowledge for biotechnological applications.
Keywords: Komagataella pastoris, nucleosome dynamics, histone structure, gene regulation, chromatin organization, X-ray crystallography, cryo-electron microscopy, biophysical assays, post-translational modifications, protein expression.