Effects of abolishing Whi2 on the proteome and nitrogen catabolite repression-sensitive protein production

0556629 - MBÚ 2023 RIV US eng J - Článek v odborném periodiku
Tate, J. - Maršíková, J. - Váchová, Libuše - Palková, Z. - Cooper, T.
Effects of abolishing Whi2 on the proteome and nitrogen catabolite repression-sensitive protein production.
G3-Genes, Genomes, Genetics. Roč. 12, č. 3 (2022), č. článku jkab432. ISSN 2160-1836. E-ISSN 2160-1836
Grant CEP: GA MŠMT(CZ) LTAUSA18162
Institucionální podpora: RVO:61388971
Klíčová slova: gata factor responses * nuclear gln3 import * rapamycin complex 1 * saccharomyces-cerevisiae * amino-acid * gene-expression * transcriptional activation * tor proteins * yeast-cells * translational control * TorC1 complex * Gln3 * Whi2 * nitrogen metabolism * synthetic complete medium * signal transduction * nuclear translocation * Gat1 * dal80 * Gcn2
Obor OECD: Microbiology
Impakt faktor: 2.6, rok: 2022
Způsob publikování: Open access
https://academic.oup.com/g3journal/article/12/3/jkab432/6468623?login=true

In yeast physiology, a commonly used reference condition for many experiments, including those involving nitrogen catabolite repression (NCR), is growth in synthetic complete (SC) medium. Four SC formulations, SCCSH,1990, SCCSH,1994, SCCSH,2005, and SCME, have been used interchangeably as the nitrogen-rich medium of choice [Cold Spring Harbor Yeast Course Manuals (SCCSH) and a formulation in the methods in enzymology (SCME)]. It has been tacitly presumed that all of these formulations support equivalent responses. However, a recent report concluded that (i) TorC1 activity is downregulated by the lower concentration of primarily leucine in SCME relative to SCCSH. (ii) The Whi2-Psr1/2 complex is responsible for this downregulation. TorC1 is a primary nitrogen-responsive regulator in yeast. Among its downstream targets is control of NCR-sensitive transcription activators Gln3 and Gat1. They in turn control production of catabolic transporters and enzymes needed to scavenge poor nitrogen sources (e.g., Proline) and activate autophagy (ATG14). One of the reporters used in Chen et al. was an NCR-sensitive DAL80-GFP promoter fusion. This intrigued us because we expected minimal if any DAL80 expression in SC medium. Therefore, we investigated the source of the Dal80-GFP production and the proteomes of wild-type and whi2 Delta cells cultured in SCCSH and SCME. We found a massive and equivalent reorientation of amino acid biosynthetic proteins in both wild-type and whi2 Delta cells even though both media contained high overall concentrations of amino acids. Gcn2 appears to play a significant regulatory role in this reorientation. NCR-sensitive DAL80 expression and overall NCR-sensitive protein production were only marginally affected by the whi2 Delta. In contrast, the levels of 58 proteins changed by an absolute value of log(2) between 3 and 8 when Whi2 was abolished relative to wild type. Surprisingly, with only two exceptions could those proteins be related in GO analyses, i.e., GO terms associated with carbohydrate metabolism and oxidative stress after shifting a whi2 Delta from SCCSH to SCME for 6 h. What was conspicuously missing were proteins related by TorC1- and NCR-associated GO terms.
Trvalý link: http://hdl.handle.net/11104/0331291