RUI: Investigating the synthesis and unique activities of bactofilins with multiple isoforms

细胞的细胞骨架在细胞的组织、运动和细胞内大分子的运动中起着重要的作用。在这个项目中，研究人员将研究细菌细胞骨架蛋白bactofilin的表达和聚合的调节。该项目将吸引具有分子微生物学培训和经验的本科生，研究细菌的细胞生物学，同时增加IONA学院的整体研究能力。本科生将接受技术培训，使他们能够将特定突变工程到非致病细菌的基因组中，并评估对细胞整体活动的影响，并为他们提供经验，以增强他们从事STEM职业的准备。这个项目的目的是为了进一步了解独特的细菌细胞骨架家族--杆菌丝蛋白--对黄色粘球菌细胞生物学的贡献。黄色微囊藻表达杆状丝蛋白的两种异构体BACM。产生这两种异构体的细胞机制，每一种异构体对黄曲霉细胞生物学的潜在独特贡献，以及多种异构体杆状丝蛋白的现象有多普遍，目前都是未知的。此外，细丝在体内形成时的极性还没有确定。为了应对这些研究挑战，PI将与本科生合作，使用聚合酶链式反应指导的突变、黄花莲中突变蛋白的表达、免疫印迹、免疫荧光显微镜和蛋白质-蛋白质相互作用研究，以产生多个细菌纤连蛋白BACM异构体，并研究每个异构体如何对黄花莲细胞生物学做出贡献。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The cytoskeleton of a cell plays an important role in the organization of the cell, its motility, and the movement of macromolecules within the cell. In this project, the researcher will investigate the regulation of expression and polymerization of the bacterial cytoskeletal protein, bactofilin. This project will engage undergraduate students with training and experience in molecular microbiology, studying the cell biology of bacteria, while increasing the overall research capacity of Iona College. Undergraduates will be trained in techniques that will allow them to engineer specific mutations into the genomes of non-pathogenic bacteria and evaluate the influence on the overall activity of the cells, and provide them with experiences that will enhance their readiness to engage in STEM careers. The goal of this project is to further our understanding of the contributions of the uniquely bacterial cytoskeleton family, bactofilins, to the cell biology of Myxococcus xanthus. Two isoforms of the bactofilin, BacM, are expressed by M. xanthus. The cellular mechanisms that produce the two isoforms, the potential unique contributions of each isoform to M. xanthus cell biology, and how widespread is the phenomenon of multiple isoforms of bactofilins are all currently unknown. In addition, the polarity of the filament as it is formed in vivo has not been established. To address these research challenges, the PI will work with undergraduates using PCR-directed mutagenesis, mutant protein expression in M. xanthus, immunoblotting, immunofluorescence microscopy, and protein-protein interaction studies, to generate multiple isoforms of the bactofilin BacM and study how each isoform contributes to M. xanthus cell biology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.