yeast functional genomic screens lead to identification of a role for a bacterial effector in innate immunity regulation酵母功能基因组屏幕导致鉴定细菌的作用在先天免疫调节效应.pdf

Yeast Functional Genomic Screens Lead to Identification of a Role for a Bacterial Effector in Innate Immunity Regulation 1 1 1 2 1 1 Roger W. Kramer , Naomi L. Slagowski , Ngozi A. Eze , Kara S. Giddings , Monica F. Morrison , Keri A. Siggers , 2 1* Michael N. Starnbach , Cammie F. Lesser 1 Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Harvard Medical School, Cambridge, Massachusetts, United States of America, 2 Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts, United States of America Numerous bacterial pathogens manipulate host cell processes to promote infection and ultimately cause disease through the action of proteins that they directly inject into host cells. Identification of the targets and molecular mechanisms of action used by these bacterial effector proteins is critical to understanding pathogenesis. We have developed a systems biological approach using the yeast Saccharomyces cerevisiae that can expedite the identification of cellular processes targeted by bacterial effector proteins. We systematically screened the viable yeast haploid deletion strain collection for mutants hypersensitive to expression of the Shigella type III effector OspF. Statistical data mining of the results identified several cellular processes, including cell wall biogenesis, which when impaired by a deletion caused yeast to be hypersensitive to OspF expression. Microarray experiments revealed that OspF expression resulted in reversed regulation of genes regulated by the yeast cell wall integrity pathway. The yeast cell wall integrity pathway is a highly conserved mitogen-activated protein kinase (MAPK) signaling path