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Dryad

Data from: Live-cell analysis of IMPDH protein levels during yeast colony growth provides insights into the regulation of GTP synthesis

Cite this dataset

Shand, Erica et al. (2024). Data from: Live-cell analysis of IMPDH protein levels during yeast colony growth provides insights into the regulation of GTP synthesis [Dataset]. Dryad. https://doi.org/10.5061/dryad.80gb5mkzb

Abstract

Here we present real-time, live-cell analysis of accumulation of the Imd2 isoform of IMPDH in Saccharomyces cervisiae yeast cells forming a monolayer colony in a microfluidic device over a 50-hour time course. We observe two distinct phases of increased Imd2 accumulation: a guanine-insensitive phase early in outgrowth and a guanine-sensitive phase later, when cells become crowded. We show that the IMPDH inhibitor mycophenolic acid enhances both phases of increase. Deletion of a transcription attenuator upstream of the mRNA start site that decreases Imd2 mRNA synthesis in the presence of high GTP increases the baseline level of Imd2 protein ten-fold and abolishes guanine-sensitive but not guanine-insensitive induction. Our results suggest that at least two mechanisms of yeast Imd2 regulation exist, the known GTP-dependent attenuation of RNA polymerase II elongation and a GTP concentration-independent pathway that may be controlled by cell growth state.

Methods

Refer associated mBio paper.

Funding

National Institute of General Medical Sciences, Award: R01GM082956

National Institute of General Medical Sciences, Award: R35GM118075

National Institute of General Medical Sciences, Award: R35GM128873

National Human Genome Research Institute, Award: T32HG002760

National Institute of General Medical Sciences, Award: T32GM08349