Expression of the atf1+ gene is upregulated in fission yeast under nitrosative and nutritional stresses

2009 ◽  
Vol 55 (11) ◽  
pp. 1323-1327 ◽  
Author(s):  
S.-H. Song ◽  
B.-M. Kim ◽  
C.-J. Lim ◽  
Y.S. Song ◽  
E.-H. Park
Keyword(s):  
Nitric Oxide ◽  
Schizosaccharomyces Pombe ◽  
Sodium Nitroprusside ◽  
Fission Yeast ◽  
Fusion Gene ◽  
Lacz Fusion ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
Nitrogen Depletion ◽  
Rt Pcr

This work was designed to assess regulation of the atf1+ gene in the fission yeast Schizosaccharomyces pombe under nitrosative and nutritional stresses, using the atf1+–lacZ fusion gene and RT–PCR. Nitric oxide (NO)-generating sodium nitroprusside (SNP; 10 µmol/L) and nitrogen depletion significantly enhanced synthesis of β-galactosidase from the atf1+–lacZ fusion gene in S. pombe Pap1-positive KP1 cells, but not in S. pombe Pap1-negative TP108-3C cells. SNP (10 µmol/L) and nitrogen depletion also caused a significant increase in atf1+ mRNA levels in Pap1-positive cells, but not in Pap1-negative cells. Depletion of glucose marginally increased synthesis of β-galactosidase from the fusion gene in S. pombe Pap1-positive cells. Taken together, the S. pombe atf1+ gene is upregulated by nitrosative and nutritional stresses on a transcriptional level, possibly via the mediation of Pap1.

scholarly journals Nitrogen depletion in the fission yeast Schizosaccharomyces pombe causes nucleosome loss in both promoters and coding regions of activated genes

2010 ◽  
Vol 20 (3) ◽  
pp. 361-371 ◽  
Author(s):  
C. Kristell ◽  
J. Orzechowski Westholm ◽  
I. Olsson ◽  
H. Ronne ◽  
J. Komorowski ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nitrogen Depletion ◽  
Coding Regions

scholarly journals Key Function for the CCAAT-Binding Factor Php4 To Regulate Gene Expression in Response to Iron Deficiency in Fission Yeast

2008 ◽  
Vol 7 (3) ◽  
pp. 493-508 ◽  
Author(s):  
Alexandre Mercier ◽  
Stephen Watt ◽  
Jürg Bähler ◽  
Simon Labbé
Keyword(s):  
Iron Deficiency ◽  
Fission Yeast ◽  
Tricarboxylic Acid Cycle ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
Gata Factor ◽  
Iron Starvation ◽  
A Genome ◽  
Binding Factor ◽  
Genes Encoding

ABSTRACT The fission yeast Schizosaccharomyces pombe responds to the deprivation of iron by inducing the expression of the php4 + gene, which encodes a negative regulatory subunit of the heteromeric CCAAT-binding factor. Once formed, the Php2/3/4/5 transcription complex is required to inactivate a subset of genes encoding iron-using proteins. Here, we used a pan-S. pombe microarray to study the transcriptional response to iron starvation and identified 86 genes that exhibit php4 + -dependent changes on a genome-wide scale. One of these genes encodes the iron-responsive transcriptional repressor Fep1, whose mRNA levels were decreased after treatment with the permeant iron chelator 2,2′-dipyridyl. In addition, several genes encoding the components of iron-dependent biochemical pathways, including the tricarboxylic acid cycle, mitochondrial respiration, amino acid biosynthesis, and oxidative stress defense, were downregulated in response to iron deficiency. Furthermore, Php4 repressed transcription when brought to a promoter using a yeast DNA-binding domain, and iron deprivation was required for this repression. On the other hand, Php4 was constitutively active when glutathione levels were depleted within the cell. Based on these and previous results, we propose that iron-dependent inactivation of Php4 is regulated at two distinct levels: first, at the transcriptional level by the iron-responsive GATA factor Fep1 and second, at the posttranscriptional level by a mechanism yet to be identified, which inhibits Php4-mediated repressive function when iron is abundant.

scholarly journals AR Deficiency Inhibits LPS-induced M1 Response in Macrophages by Activating Autophagy

2020 ◽  
Author(s):  
Peng Cheng ◽  
Jianwei Xie ◽  
Zhiyong Liu ◽  
Jian Wang
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inflammatory Responses ◽  
Macrophage Polarization ◽  
Critical Role ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
M1 Polarization ◽  
M1 Macrophage ◽  
Oxide Synthase

Abstract Macrophage M1 polarization mediates inflammatory responses and tissue damage. Recently, aldose reductase (AR) has been shown to play a critical role in of M1 polarization in macrophages. However, the underlying mechanisms are unknown. Here, we demonstrated, for the first time, that AR deficiency repressed the induction of inducible nitric oxide synthase in lipopolysaccharide (LPS)-stimulated macrophages via activation of autophagy. This suppression was related to a defect in the inhibitor of nuclear factor κB (NF-κB) kinase (IKK) complex in the classical NF-κB pathway. However, the mRNA levels of the IKKβ and IKKγ were not reduced in LPS-treated AR knockout (KO) macrophages, indicating that their proteins were downregulated at the post-transcriptional level. We discovered that LPS stimuli induced the recruitment of more beclin1 and increased autophagosome formation in AR-deficient macrophages. Blocking autophagy by 3-methyladenine and ammonium chloride treatment restored IKKβ and IKKγ protein levels and increased nitric oxide synthase production in LPS-stimulated AR-deficient macrophages. More assembled IKKβ and IKKγ undergo ubiquitination and recruit the autophagic adaptor p62 in LPS-induced AR KO macrophages, promoting their delivery to autophagosomes and lysosomes. Collectively, these findings suggest that AR deficiency involves in the regulation of NF-κB signaling, and extends the role of selective autophagy in fine-tuned M1 macrophage polarization.

Stress-dependent regulation of a monothiol glutaredoxin gene from Schizosaccharomyces pombe

2005 ◽  
Vol 51 (7) ◽  
pp. 613-620 ◽  
Author(s):  
Hong-Gyum Kim ◽  
Byung-Chul Kim ◽  
Eun-Hee Park ◽  
Chang-Jin Lim
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Potassium Chloride ◽  
Sodium Nitroprusside ◽  
Fusion Gene ◽  
Mrna Level ◽  
Carbon Sources ◽  
Lacz Gene ◽  
Gene Encoding ◽  
Stress Dependent

Glutaredoxin (Grx) is a small, heat-stable protein acting as a multi-functional glutathione-dependent disulfide oxidoreductase. In this work, a gene encoding the monothiol glutaredoxin Grx4 was cloned from the genomic DNA of the fission yeast Schizosaccharomyces pombe. The determined DNA sequence carries 1706 bp, which is able to encode the putative 244 amino acid sequence of Grx with 27 099 Da. It does not contain an intron, and the sequence CGFS is found in the active site. Grx activity was increased 1.46-fold in S. pombe cells harboring the cloned Grx4 gene, indicating that the Grx4 gene is in vivo functioning. Although aluminum, cadmium, and hydrogen peroxide marginally enhanced the synthesis of β-galactosidase from the Grx4-lacZ fusion gene, NO-generating sodium nitroprusside (0.5 mmol/L and 1.0 mmol/L) and potassium chloride (0.2 mol/L and 0.5 mol/L) significantly enhanced it. The Grx4 mRNA level was also enhanced after the treatment with sodium nitroprusside and potassium chloride. The synthesis of β-galactosidase from the Grx4-lacZ gene was increased by fermentable carbon sources, such as glucose (lower than 2%) and sucrose, but not by nonfermentable carbon sources such as acetate and ethanol. The basal expression of the S. pombe Grx4 gene did not depend on the presence of Pap1. These results imply that the S. pombe monothiol Grx4 gene is genuinely functional and regulated by a variety of stresses.Key words: monothiol glutaredoxin, Pap1, regulation, Schizosaccharomyces pombe, stress response.

scholarly journals Aldose reductase deficiency inhibits LPS-induced M1 response in macrophages by activating autophagy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Peng Cheng ◽  
Jianwei Xie ◽  
Zhiyong Liu ◽  
Jian Wang
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Aldose Reductase ◽  
Macrophage Polarization ◽  
Critical Role ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
M1 Polarization ◽  
M1 Macrophage ◽  
Oxide Synthase

AbstractMacrophage M1 polarization mediates inflammatory responses and tissue damage. Recently, aldose reductase (AR) has been shown to play a critical role in M1 polarization in macrophages. However, the underlying mechanisms are unknown. Here, we demonstrated, for the first time, that AR deficiency repressed the induction of inducible nitric oxide synthase in lipopolysaccharide (LPS)-stimulated macrophages via activation of autophagy. This suppression was related to a defect in the inhibitor of nuclear factor κB (NF-κB) kinase (IKK) complex in the classical NF-κB pathway. However, the mRNA levels of IKKβ and IKKγ were not reduced in LPS-treated AR knockout (KO) macrophages, indicating that their proteins were downregulated at the post-transcriptional level. We discovered that LPS stimuli induced the recruitment of more beclin1 and increased autophagosome formation in AR-deficient macrophages. Blocking autophagy through 3-methyladenine and ammonium chloride treatment restored IKKβ and IKKγ protein levels and increased nitric oxide synthase production in LPS-stimulated AR-deficient macrophages. More assembled IKKβ and IKKγ underwent ubiquitination and recruited the autophagic adaptor p62 in LPS-induced AR KO macrophages, promoting their delivery to autophagosomes and lysosomes. Collectively, these findings suggest that AR deficiency is involved in the regulation of NF-κB signaling, and extends the role of selective autophagy in fine-tuned M1 macrophage polarization.

C/EBPα and C/EBPε Induce Monocytic Differentiation of Myelomonocytic Cells with MLL Chimeric Fusion Gene.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1938-1938
Author(s):  
Hiromichi Matsushita ◽  
Hideaki Nakajima ◽  
Yoshihiko Nakamura ◽  
Hideo Tsukamoto ◽  
Yumiko Tanaka ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Growth Inhibition ◽  
Cellular Proliferation ◽  
Fusion Gene ◽  
Flow Cytometric Analysis ◽  
Transcriptional Level ◽  
Mrna Levels ◽  
Fusion Genes ◽  
Monocytic Differentiation ◽  
Myelomonocytic Leukemia

Abstract Mixed lineage leukemia (MLL) gene, located on chromosome segment 11q23, is involved in various types of hematological malignancies including infant and secondary leukemias. To date, more than 40 partners have been reported in the recurrent translocations with 11q23, and the resultant fusion genes are believed to contribute to leukeomogenesis. In acute myeloid leukemia (AML), the incidence of MLL rearrangements is higher in subtypes with monocytic components, namely M4, M5a and M5b. Furthermore, several MLL fusion genes are known to cause differentiation arrest at the myelo-monocytic stage when they are expressed in murine hematopietic cells. Recent studies have revealed that the myeloid transcription factors, CCAAT/enhancer binding proteins (C/EBPs), regulate not only granulocytic but also monocytic differentiation. Accordingly, we investigated the roles of C/EBPα and C/EBPε in myelomonocytic leukemia with MLL fusion genes. The protein expression of C/EBPα and C/EBPε was induced in retinoic acid-mediated monocytic differentiation of human AML cell lines with MLL-AF9, THP-1 and MOLM-14. Furthermore, 9-cis Retinoic acid upregulated the mRNA levels of Cebpa and Cebpe in monocytic differentiation of an IL-3-dependent murine cell line carrying MLL-SEPT6, HF-6. When HF-6 cells were retrovirally infected with inducible forms of C/EBPα (C/EBPα-ER) or C/EBPε (C/EBPε-ER), mature monocytes were markedly induced. Flow cytometric analysis revealed the increased intensity of Mac-1 and Gr-1 with C/EBPα or C/EBPε induction. In addition, their cell growth was severely impaired. This growth inhibition was believed to be partly due to induction of apoptosis. There were no apparent differences in monocytic differentiation induction and growth inhibition between C/EBPα and C/EBPε. The overexpression of C/EBPα or C/EBPε in HF-6 cells was found to repress the gene expressions of Myc, Hoxa7 and Hoxa9, and induced CdknIa and SfpiI in quantitative RT-PCR analysis. The transcriptional level of C/EBPε was up-regulated by the induction of C/EBPα, but not vice versa. Since FLT3-ITD has been detected in 20 to 30% of AML cases and is known to cause a more immature type of AML with MLL-SEPT6 in vivo analysis, we subsequently introduced internal tandem duplication of FLT3 (FLT3-ITD) into HF-6 cells. These cells could proliferate without IL-3. Overexpression of C/EBPα and C/EBPε in these cells, however, inhibited the cellular proliferation and induced monocytic differentiation, as was seen in the original HF-6 cells. MOLM-14 cells, also possessing FLT3-ITD, have recently reported to differentiate by overexpression of C/EBPα (J Exp Med2006: 203: 371–381). Taken together, these findings suggest that not only C/EBPα but C/EBPε alone can induce monocytic differentiation of myelomonocytic leukemia cells with MLL fusion gene, regardless of the existence of FLT3-ITD, and that this process is accompanied by down-regulation of the genes which are related to MLL-induced AML leukemogenesis.

Overexpression of a metacaspase gene stimulates cell growth and stress response inSchizosaccharomyces pombe

2007 ◽  
Vol 53 (8) ◽  
pp. 1016-1023 ◽  
Author(s):  
Hye-Won Lim ◽  
Su-Jung Kim ◽  
Eun-Hee Park ◽  
Chang-Jin Lim
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Stress Response ◽  
Cell Growth ◽  
Schizosaccharomyces Pombe ◽  
Sodium Nitroprusside ◽  
Fission Yeast ◽  
Recombinant Plasmid ◽  
Mrna Level ◽  
Yeast Cells ◽  
Yeast Saccharomyces Cerevisiae

A unique gene named pca1+, encoding a metacaspase, was cloned from the fission yeast Schizosaccharomyces pombe and was used to create a recombinant plasmid, pPMC. The metacaspase mRNA level was markedly elevated in the fission yeast cells harboring the plasmid pPMC. Overexpressed Pca1+appeared to stimulate the growth of the fission yeast cells instead of arresting their growth. Its expression was enhanced by stress-inducing agents such as H2O2, sodium nitroprusside, and CdCl2, and it conferred cytoprotection, especially against CdCl2. However, such protection was not reproducible in the budding yeast Saccharomyces cerevisiae harboring pPMC. Taken together, these results propose that Pca1+may be involved in the growth and stress response of the fission yeast.

Stress-dependent regulation of Pbh1, a BIR domain-containing protein, in the fission yeast

2006 ◽  
Vol 52 (12) ◽  
pp. 1261-1265 ◽  
Author(s):  
Nam-Chul Cho ◽  
Hyun-Jung Kang ◽  
Hye-Won Lim ◽  
Byung-Chul Kim ◽  
Eun-Hee Park ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nitrosative Stress ◽  
Nitrogen Starvation ◽  
Fusion Gene ◽  
Shuttle Vector ◽  
Upstream Region ◽  
Basal Expression ◽  
Stress Dependent ◽  
Bir Domain

To elicit the physiological roles of Pbh1, a baculoviral IAP repeat (BIR) domain-containing protein, in Schizosaccharomyces pombe, we investigated if Pbh1 expression is regulated by stress. The upstream region (1221 bp) of the pbh1 gene was fused into the promoterless β-galactosidase gene of the shuttle vector YEp367R, and the resultant fusion plasmid was named pPbh04. The synthesis of β-galactosidase from the pbh1-lacZ fusion gene was markedly enhanced by sodium nitroprusside (SNP) generating nitric oxide. The basal expression of the pbh1 gene required the presence of Pap1. Pap1 also mediated the induction of the pbh1 gene by SNP and nitrogen starvation. Pap1-dependent induction of the pbh1 gene by SNP was confirmed by the enhanced level of the pbh1 mRNA in Pap1-positive cells but not in Pap1-negative cells. Taken together, it was demonstrated that the pbh1 genes are positively regulated by nitrosative and nitrogen starvation stresses in Pap1-dependent manner.Key words: fission yeast, nitrosative stress, nutritional stress, nitrogen starvation, Pap1, Pbh1, regulation, Schizosaccharomyces pombe.

Sign in / Sign up

Export Citation Format

Share Document