Transcriptional and post-translational regulation of neutral trehalase in Schizosaccharomyces pombe during thermal stress.
In the fission yeast Schizosaccharomyces pombe, a heat shock enhances transcription of the ntp1(+) gene, encoding the hydrolytic enzyme neutral trehalase. As compared to wild-type cells, cells devoid of the MAP kinase Sty1p showed a strong decrease in ntp1(+) expression induced by the temperature upshift, indicating that the stress-activated protein kinase (SAPK) pathway regulates the expression of this gene during heat shock. The transcription factor Atf1p, which is the main downstream target for Sty1p in the SAPK pathway, appears to be involved in such control, since ntp1(+) expression under heat shock proved to be significantly blocked in atf1(+)-disrupted cells. Serial deletion and point mutation analyses of the ntp1(+) promoter, as well as electrophoretic mobility shift assays, revealed the existence of a CRE-like element as the target for Atf1p-mediated expression under thermal stress. The relevance of two putative HSE elements located in the ntp1(+) promoter was also investigated for their potential role in regulating ntp1(+) transcription during heat shock. The results support a model in which heat-induced Atf1p binding to the CRE-like element favours the subsequent interaction of the heat shock factor (HSF) with HSE elements in the ntp1(+) promoter. Unlike what happens under osmostress or oxidative treatments, Sty1p has no role in the post-translational activation of neutral trehalase induced by heat shock in the fission yeast.
Paredes
Vanessa
V
Department of Genetics and Microbiology, Facultad de Biología, University of Murcia, 30071 Murcia, Spain.
Franco
Alejandro
A
Madrid
Marisa
M
Soto
Teresa
T
Vicente-Soler
Jero
J
Gacto
Mariano
M
Cansado
José
J
D051697:Activating Transcription Factor 1; D015152:Blotting, Northern; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D015966:Gene Expression Regulation, Fungal; D018869:Heat-Shock Response / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000235:genetics / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism*; D015533:Transcriptional Activation
D051697:Activating Transcription Factor 1; D004271:DNA, Fungal; D010750:Phosphoproteins; D012331:RNA, Fungal; D029702:Schizosaccharomyces pombe Proteins; C106740:atf1 protein, S pombe; D020928:Mitogen-Activated Protein Kinases; C095409:sty1 protein, S pombe; C464880:Ntp1p protein, S pombe; D010744:Phosphoric Monoester Hydrolases