Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.1 (protein kinase)
 81,284 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cells respond to increased external osmolarities by enhanced accumulation of compatible solutes. In yeast-cells, mainly exemplified by Saccharomyces cerevisiae, the premier compatible solute is the polyhydroxy-alcohol glycerol, the production of which is accompanied by overall metabolic changes. By applying two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) coupled to computerized image quantification, a large body of valuable physiological information relating to this stress-adaptation has been gathered. One of the presumed key-enzymes in the production of glycerol in the cell is glycerol 3-phosphate dehydrogenase encoded by the GPD1 gene. The amount of this protein is enhanced during saline stress, and from 2-D analysis linked to microsequencing it became apparent that the osmo-regulated from contained a putative presequence. Sequence analysis of another salt-induced spot in the 2-D pattern revealed identity to a gene, YER062c, with previously unknown function. Biochemical characterization of this protein, including standard purification via chromatography and subsequent activity/specificity measurements, identified this salt-regulated protein as the missing protein/gene in glycerol production, namely the glycerol 3-phosphatase. The sequence of another salt regulated protein resolved in the 2-D gel revealed identity to a bacterial dihydroxyacetone kinase, thus indicating salt induced glycerol dissimilation. Comparing Northern data to the 2-D generated expression pattern revealed a strong correlation, indicating mainly regulation at the transcriptional level. In addition, altered expression during saline growth of some of the glycolytic enzymes was also apparent. Signalling mutants, either in the cAMP-dependent protein kinase A pathway or in a protein kinase cascade, have been analyzed during osmotic stress via 2-D PAGE, grouping proteins/genes apparently regulated via similar mechanismus. Proteome analysis has proven invaluable in the unravelling of the molecular physiology of yeast cells during adaptation and growth under osmotic stress, identifying vital components not selected by purely genetic approaches.
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PMID:Osmoresponsive proteins and functional assessment strategies in Saccharomyces cerevisiae. 929 57

We have characterized the strongly homologous GPP1/RHR2 and GPP2/HOR2 genes, encoding isoforms of glycerol 3-phosphatase. Mutants lacking both GPP1 and GPP2 are devoid of glycerol 3-phosphatase activity and produce only a small amount of glycerol, confirming the essential role for this enzyme in glycerol biosynthesis. Overproduction of Gpp1p and Gpp2p did not significantly enhance glycerol production, indicating that glycerol phosphatase is not rate-limiting for glycerol production. Previous studies have shown that expression of both GPP1 and GPP2 is induced under hyperosmotic stress and that induction partially depends on the HOG (high osmolarity glycerol) pathway. We here show that expression of GPP1 is strongly decreased in strains having low protein kinase A activity, although it is still responsive to osmotic stress. The gpp1Delta/gpp2Delta double mutant is hypersensitive to high osmolarity, whereas the single mutants remain unaffected, indicating GPP1 and GPP2 substitute well for each other. Transfer to anaerobic conditions does not affect expression of GPP2, whereas GPP1 is transiently induced, and mutants lacking GPP1 show poor anaerobic growth. All gpp mutants show increased levels of glycerol 3-phosphate, which is especially pronounced when gpp1Delta and gpp1Delta/gpp2Delta mutants are transferred to anaerobic conditions. The addition of acetaldehyde, a strong oxidizer of NADH, leads to decreased glycerol 3-phosphate levels and restored anaerobic growth of the gpp1Delta/gpp2Delta mutant, indicating that the anaerobic accumulation of NADH causes glycerol 3-phosphate to reach growth-inhibiting levels. We also found the gpp1Delta/gpp2Delta mutant is hypersensitive to the superoxide anion generator, paraquat. Consistent with a role for glycerol 3-phosphatase in protection against oxidative stress, expression of GPP2 is induced in the presence of paraquat. This induction was only marginally affected by the general stress-response transcriptional factors Msn2p/4p or protein kinase A activity. We conclude that glycerol metabolism plays multiple roles in yeast adaptation to altered growth conditions, explaining the complex regulation of glycerol biosynthesis genes.
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PMID:The yeast glycerol 3-phosphatases Gpp1p and Gpp2p are required for glycerol biosynthesis and differentially involved in the cellular responses to osmotic, anaerobic, and oxidative stress. 1105 91