Investigating cell autonomy in microorganisms

• Published in: Current genetics Vol. 68; no. 2; pp. 305 - 318
• Main Authors: Piccirillo, Sarah, Morgan, Andrew P., Leon, Andy Y., Smith, Annika L., Honigberg, Saul M.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.04.2022; Springer Nature B.V
• Subjects: Assaying; Autonomy; Biochemistry; Biomedical and Life Sciences; Catabolite repression; Cell Biology; Cell differentiation; Cell signaling; Codon, Terminator; Colonies; Differentiation (biology); E coli; Gene silencing; Genetic effects; Life Sciences; Localization; Microbial Genetics and Genomics; Microbiology; Microorganisms; Plant Sciences; Protein Biosynthesis; Proteomics; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Signaling; Technical Note; Yeast; Yeasts; Colony patterning; Density dependence; Cell–cell signal; Localization; Cell-nonautonomy; Escherichia coli; Saccharomyces cerevisiae
• ISSN: 0172-8083; 1432-0983
• DOI: 10.1007/s00294-022-01231-5

Cell–cell signaling in microorganisms is still poorly characterized. In this Methods paper, we describe a genetic procedure for detecting cell-nonautonomous genetic effects, and in particular cell–cell signaling, termed the chimeric colony assay (CCA). The CCA measures the effect of a gene on a biological response in a neighboring cell. This assay can measure cell autonomy for range of biological activities including transcript or protein accumulation, subcellular localization, and cell differentiation. To date, the CCA has been used exclusively to investigate colony patterning in the budding yeast Saccharomyces cerevisiae . To demonstrate the wider potential of the assay, we applied this assay to two other systems: the effect of Grr1 on glucose repression of GAL1 transcription in yeast and the effect of rpsL on stop-codon translational readthrough in Escherichia coli . We also describe variations of the standard CCA that address specific aspects of cell–cell signaling, and we delineate essential controls for this assay. Finally, we discuss complementary approaches to the CCA. Taken together, this Methods paper demonstrates how genetic assays can reveal and explore the roles of cell–cell signaling in microbial processes.