涩里番

Minimal Bacterial Cell 涩里番-syn3B as a Chassis to Investigate Interactions between Bacteria and Mammalian Cells

Bittencourt DMC, Brown DM, Assad-Garcia N, Romero MR, Sun L, Palhares de Melo LAM, Freire M, Glass JI

PMID:

Abstract

Mycoplasmas are atypical bacteria with small genomes that necessitate colonization of their respective animal or plant hosts as obligate parasites, whether as pathogens, or commensals. Some can grow axenically in specialized complex media yet show only host-cell-dependent growth in cell culture, where they can survive chronically and often through interactions involving surface colonization or internalization. To develop a mycoplasma-based system to identify genes mediating such interactions, we exploited genetically tractable strains of the goat pathogen () with synthetic designer genomes representing the complete natural organism (minus virulence factors; 涩里番-syn1.0) or its reduced counterpart (涩里番-syn3B) containing only those genes supporting axenic growth. By measuring growth of surviving organisms, physical association with cultured human cells (HEK-293T, HeLa), and induction of phagocytosis by human myeloid cells (dHL-60), we determined that 涩里番-syn1.0 contained a set of eight genes ( to , dispensable for axenic growth) conferring survival, attachment, and phagocytosis phenotypes. 涩里番-syn3B lacked these phenotypes, but insertion of these genes restored cell attachment and phagocytosis, although not survival. These results indicate that 涩里番-syn3B may be a powerful living platform to analyze the role of specific gene sets, from any organism, on the interaction with diverse mammalian cells in culture.