Dataset Information

Proteomic consequences of TDA1 deficiency in Saccharomyces cerevisiae — Protein kinase Tda1 is essential for Hxk1 and Hxk2 serine 15 phosphorylation

ABSTRACT: Hexokinase 2 (Hxk2) of Saccharomyces cerevisiae is a dual function hexokinase, acting as a glycolytic enzyme and being involved in the transcriptional regulation of glucose-repressible genes. Relief from glucose repression is accompanied by phosphorylation of Hxk2 at serine 15, which has been attributed to the protein kinase Tda1. To explore the role of Tda1 beyond Hxk2 phosphorylation the proteomic consequences of TDA1 deficiency were investigated by difference gel electrophoresis (2D-DIGE) comparing a wild type and a Δtda1 deletion mutant. Both were grown at 2 % and 0.1 % (w/v) glucose to additionally regard glucose repression/derepression. A total of eight spots exhibiting a minimum 2-fold enhanced or reduced fluorescence upon TDA1 deficiency was detected and identified by mass spectrometry. These include – besides the expected Hxk2 – two proteoforms of hexokinase 1 (Hxk1). Targeted proteomics analyses in conjunction with 2D-DIGE demonstrated that TDA1 is indispensable for Hxk2 and Hxk1 phosphorylation at serine 15. 35 glucose-dependent proteins were identified. To improve spot quantification, a simple method approximating spots as rotationally symmetric solids is presented along with new data on the quantities of Hxk1 and Hxk2 and their serine 15 phosphorylated forms at high and low glucose growth conditions. The Δtda1 deletion mutant exhibited no altered growth comparing high or low glucose conditions or on alternative carbon sources. Also, invertase activity, serving as a reporter for glucose derepression, was not significantly altered. Instead, an involvement of Tda1 in oxidative stress response is suggested.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture

SUBMITTER: Marc Gentzel

LAB HEAD: Dr. Karina Kettner

PROVIDER: PXD030105 | Pride | 2023-02-06

REPOSITORIES: Pride

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			Action	DRS
	20180629_TK01_01_RTSTD_16fm_45min_01_180629183024.mgf	Mgf
	20180629_TK01_01_RTSTD_16fm_45min_01_180629183024.raw	Raw
	20180629_TK01_01_RTSTD_16fm_45min_01_180629183024_F005047.dat	Other
	20180629_TK01_02_RTSTD_16fm_45min_01_180629200901.mgf	Mgf
	20180629_TK01_02_RTSTD_16fm_45min_01_180629200901.raw	Raw

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Proteomic consequences of TDA1 deficiency in Saccharomyces cerevisiae: Protein kinase Tda1 is essential for Hxk1 and Hxk2 serine 15 phosphorylation.

Müller Henry H Lesur Antoine A Dittmar Gunnar G Gentzel Marc M Kettner Karina K

Scientific reports 20221027 1

Hexokinase 2 (Hxk2) of Saccharomyces cerevisiae is a dual function hexokinase, acting as a glycolytic enzyme and being involved in the transcriptional regulation of glucose-repressible genes. Relief from glucose repression is accompanied by phosphorylation of Hxk2 at serine 15, which has been attributed to the protein kinase Tda1. To explore the role of Tda1 beyond Hxk2 phosphorylation, the proteomic consequences of TDA1 deficiency were investigated by difference gel electrophoresis (2D-DIGE) co ...[more]

PMID: 36302925

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