Detection of cyclosporin A and its cytosolic binding protein, cyclophilin, in CYC A high-producing strain of Tolypocladium terricola

1992 ◽  
Vol 50 (1) ◽  
pp. 790-791
Author(s):  
Vladimír Matha ◽  
Alexander Jegorov ◽  
Frantíšek Weyda
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Bone Marrow ◽  
Cyclosporin A ◽  
Binding Protein ◽  
Human Medicine ◽  
Antifungal Compound ◽  
Primary Target ◽  
Kidney Liver ◽  
Transplanted Organs

Cyclosporin A (Cyc A), a cyclic fungal undecapeptide produced by various deuteromyces is a potent immunosuppressant widely used in human medicine to prevent rejection of transplanted organs such a kidney, liver, heart and bone marrow. Although originally isolated as an antifungal compound, practically no attention was devoted to its effect on the physiology of its high-producing organisms. In vertebrate cells, the cytosolic Cyc-A binding protein, cyclophilin, is expected to be a primary target involved in the regulation of Cyc A immunosuppressive effect. Similarly, the direct cyclophilin-mediated effect (toxicity) confirming the key role of this Cyc A receptor in Cyc A function, was determined even in Neurospora crassa and Saccharomyces cerevisiae. Induction of Cyc A resistance in both microorganisms was characterized by either of the complete lost of cyclophilin or by the lost of its ability to bind Cyc A.

scholarly journals The Saccharomyces cerevisiae SPT8 gene encodes a very acidic protein that is functionally related to SPT3 and TATA-binding protein.

Genetics ◽  
1994 ◽  
Vol 137 (3) ◽  
pp. 647-657 ◽  
Author(s):  
D M Eisenmann ◽  
C Chapon ◽  
S M Roberts ◽  
C Dollard ◽  
F Winston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Protein ◽  
Tata Binding Protein ◽  
Sequence Motif ◽  
Heterotrimeric G Proteins ◽  
Eukaryotic Proteins ◽  
Multiple Copies ◽  
The Relationship ◽  
Insertion Mutations

Abstract Mutations in the Saccharomyces cerevisiae SPT8 gene were previously isolated as suppressors of retrotransposon insertion mutations in the 5' regions of the HIS4 and LYS2 genes. Mutations in SPT8 confer phenotypes similar to those caused by particular mutations in SPT15, which encodes the TATA-binding protein (TBP). These phenotypes are also similar to those caused by mutations in the SPT3 gene, which encodes a protein that directly interacts with TBP. We have now cloned and sequenced the SPT8 gene and have shown that it encodes a predicted protein of 602 amino acids with an extremely acidic amino terminus. In addition, the predicted SPT8 amino acid sequence contains one copy of a sequence motif found in multiple copies in a number of other eukaryotic proteins, including the beta subunit of heterotrimeric G proteins. To investigate further the relationship between SPT8, SPT3 and TBP, we have analyzed the effect of an spt8 null mutation in combination with different spt3 and spt15 mutations. This genetic analysis has shown that an spt8 deletion mutation is suppressed by particular spt3 alleles. Taken together with previous results, these data suggest that the SPT8 protein is required, directly or indirectly, for TBP function at particular promoters and that the role of SPT8 may be to promote a functional interaction between SPT3 and TBP.

scholarly journals Impaired erythropoietin production in mice treated with cyclosporin A

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1615-1618 ◽  
Author(s):  
AM Vannucchi ◽  
A Grossi ◽  
A Bosi ◽  
D Rafanelli ◽  
S Guidi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Animal Model ◽  
Cyclosporin A ◽  
Renal Toxicity ◽  
Reference Population ◽  
Marrow Transplant ◽  
Allogeneic Bone ◽  
Kidney Toxicity ◽  
Erythropoietin Production

Abstract Because recent data indicate that erythropoietin (Epo) production is defective in allogeneic bone marrow transplant (BMT) patients, we investigated the role of the immunosuppressant, nephrotoxic, agent cyclosporin A (CsA) on renal Epo production using an animal model. Mice were injected with 1.0 to 40.0 mg/kg/d CsA for 15 days. Thereafter, circulating Epo levels were evaluated in both intact animals and in mice made anemic with phenylhydrazine (PHZ). Serum Epo levels measured in CsA-treated animals were then compared with the predicted levels, which had been calculated in a reference population of normal, either intact or anemic, mice. In CsA-treated, intact animals both hematocrit and serum Epo levels were not significantly different from controls. However, serum Epo levels in CsA-treated, anemic mice were significantly lower than those expected in a control population of untreated, anemic mice with similar degrees of anemia. No significant increase in serum creatinine was recorded even at the highest doses of CsA used, nor were we able to document signs of renal toxicity by histologic examination of the kidneys. Therefore, therapeutical doses of CsA appear to affect the production of Epo under conditions in which the demand of the hormone is increased, as in response to anemia. We suggest that a subclinical kidney toxicity produced by CsA might have a role in the pathogenesis of the impaired Epo production observed in BMT patients, and may contribute to a delayed erythroid engraftment in at least some BMT patients.

scholarly journals The Role of Cold Inducible RNA-Binding Protein in Cardiac Physiology and Diseases

2021 ◽  
Vol 12 ◽  
Author(s):  
Peng Zhong ◽  
Jianye Peng ◽  
Zhouyan Bian ◽  
He Huang
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Stress Conditions ◽  
Telomere Maintenance ◽  
Transcriptional Level ◽  
Cardiac Physiology ◽  
Electrophysiological Properties ◽  
Kidney Liver

Cold-inducible RNA-binding protein (CIRP) is an intracellular stress-response protein that can respond to various stress conditions by changing its expression and regulating mRNA stability. As an RNA-binding protein, CIRP modulates gene expression at the post-transcriptional level, including those genes involved in DNA repair, cellular redox metabolism, circadian rhythms, telomere maintenance, and cell survival. CIRP is expressed in a large variety of tissues, including testis, brain, lung, kidney, liver, stomach, bone marrow, and heart. Recent studies have observed the important role of CIRP in cardiac physiology and diseases. CIRP regulates cardiac electrophysiological properties such as the repolarization of cardiomyocytes, the susceptibility of atrial fibrillation, and the function of the sinoatrial node in response to stress. CIRP has also been suggested to protect cardiomyocytes from apoptosis under various stress conditions, including heart failure, high glucose conditions, as well as during extended heart preservation under hypothermic conditions. This review summarizes the findings of CIRP investigations in cardiac physiology and diseases and the underlying molecular mechanism.

The Y-box Binding Protein YB-1 Is Associated with Progressive Disease and Mediates Survival and Drug Resistance in Multiple Myeloma.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3385-3385
Author(s):  
Manik Chatterjee ◽  
Christoph Ransco ◽  
Thorsten Stühmer ◽  
Niels Eckstein ◽  
Hans-Dieter Royer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Drug Resistance ◽  
Cell Lines ◽  
Plasma Cells ◽  
Apoptotic Cell ◽  
Binding Protein ◽  
Regulation Of Transcription ◽  
Drug Induced ◽  
Bone Marrow Biopsies

Abstract Introduction: The Y-box binding protein YB-1 is a member of the cold shock domain protein superfamily and represents one of the most evolutionary conserved nucleic-acid binding proteins. Yb-1 is involved in a wide variety of cellular functions, such as regulation of transcription and translation, but also in DNA-repair and stress response to extracellular signals. Furthermore, recent reports from our group could show that overexpressed YB-1 plays a role in drug resistance of breast cancer cells and might act as an oncogene. The goal of this study was to investigate a potential pathogenetic role of YB-1 in MM. Material and Methods: For the detection of YB-1 expression in vivo, bone marrow biopsies of MM patients were analyzed by immunohistochemistry. The expression of YB-1 protein in a number of MM cell lines was analyzed by Western Blotting. The regulation of YB-1 expression through major signaling pathways, e.g. IL-6R/STAT3, Ras/MAPK and PI3K/AKT, was analyzed using specific inhibitors of these pathways (Sant7, PD98059 and Ly294002). To determine the role of YB-1 for survival and proliferation, siRNA technology was exploited to transiently knockdown the expression of YB-1 in the MM cell lines INA-6 and MM.1s. In addition, these YB-1 knockdown cells were exposed to doxorubicin and melphalan to evaluate the influence of YB-1 on drug resistance. Results: Immunohistochemical analyses of bone marrow biopsies revealed that YB-1 is strongly expressed only in MM cells of samples that show a highly proliferative phenotype. This subgroup of MM patients was characterized by an aggressive clinical course. In contrast, MM cells of samples with a slow proliferative status did not show YB-1 expression. Interestingly, tumor cells of patients that responded to chemotherapy did not express YB-1. Furthermore, neither normal bone marrow plasma cells nor premalignant plasma cells of MGUS patients showed YB-1 expression. YB-1 was detected in all of the evaluated MM cell lines. YB-1 expression was not regulated by the IL-6R/STAT3, Ras/MAPK and PI3K/AKT pathways. Knockdown of YB-1 in INA-6 and MM1.s cells by siRNA resulted in a slow proliferation phenotype with a higher apoptotic cell fraction. In addition, we observed that YB-1 knockdown remarkably sensitized cells towards drug-induced apoptosis. Conclusions: YB-1 expression in MM appears to be correlated with a highly proliferative phenotype and with disease progression. YB-1 contributes to the malignant growth and drug resistance and might be therefore an attractive therapeutical target to circumvent aquired drug resistance in MM.

scholarly journals Impaired erythropoietin production in mice treated with cyclosporin A

Blood ◽  
1991 ◽  
Vol 78 (6) ◽  
pp. 1615-1618 ◽  
Author(s):  
AM Vannucchi ◽  
A Grossi ◽  
A Bosi ◽  
D Rafanelli ◽  
S Guidi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Animal Model ◽  
Cyclosporin A ◽  
Renal Toxicity ◽  
Reference Population ◽  
Marrow Transplant ◽  
Allogeneic Bone ◽  
Kidney Toxicity ◽  
Erythropoietin Production

Because recent data indicate that erythropoietin (Epo) production is defective in allogeneic bone marrow transplant (BMT) patients, we investigated the role of the immunosuppressant, nephrotoxic, agent cyclosporin A (CsA) on renal Epo production using an animal model. Mice were injected with 1.0 to 40.0 mg/kg/d CsA for 15 days. Thereafter, circulating Epo levels were evaluated in both intact animals and in mice made anemic with phenylhydrazine (PHZ). Serum Epo levels measured in CsA-treated animals were then compared with the predicted levels, which had been calculated in a reference population of normal, either intact or anemic, mice. In CsA-treated, intact animals both hematocrit and serum Epo levels were not significantly different from controls. However, serum Epo levels in CsA-treated, anemic mice were significantly lower than those expected in a control population of untreated, anemic mice with similar degrees of anemia. No significant increase in serum creatinine was recorded even at the highest doses of CsA used, nor were we able to document signs of renal toxicity by histologic examination of the kidneys. Therefore, therapeutical doses of CsA appear to affect the production of Epo under conditions in which the demand of the hormone is increased, as in response to anemia. We suggest that a subclinical kidney toxicity produced by CsA might have a role in the pathogenesis of the impaired Epo production observed in BMT patients, and may contribute to a delayed erythroid engraftment in at least some BMT patients.

scholarly journals Pbp1p, a Factor Interacting withSaccharomyces cerevisiae Poly(A)-Binding Protein, Regulates Polyadenylation

1998 ◽  
Vol 18 (12) ◽  
pp. 7383-7396 ◽  
Author(s):  
David A. Mangus ◽  
Nadia Amrani ◽  
Allan Jacobson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Role ◽  
Binding Protein ◽  
Carboxy Terminus ◽  
Ribosomal Subunits ◽  
Initiation Of Translation ◽  
Genes Encoding ◽  
Key Factor ◽  
Two Hybrid

ABSTRACT The poly(A) tail of an mRNA is believed to influence the initiation of translation, and the rate at which the poly(A) tail is removed is thought to determine how fast an mRNA is degraded. One key factor associated with this 3′-end structure is the poly(A)-binding protein (Pab1p) encoded by the PAB1 gene inSaccharomyces cerevisiae. In an effort to learn more about the functional role of this protein, we used a two-hybrid screen to determine the factor(s) with which it interacts. We identified five genes encoding factors that specifically interact with the carboxy terminus of Pab1p. Of a total of 44 specific clones identified,PBP1 (for Pab1p-binding protein) was isolated 38 times. Of the putative interacting genes examined, PBP1 promoted the highest level of resistance to 3-aminotriazole (>100 mM) in constructs in which HIS3 was used as a reporter. We determined that a fraction of Pbp1p cosediments with polysomes in sucrose gradients and that its distribution is very similar to that of Pab1p. Disruption ofPBP1 showed that it is not essential for viability but can suppress the lethality associated with a PAB1 deletion. The suppression of pab1Δ by pbp1Δ appears to be different from that mediated by other pab1 suppressors, since disruption of PBP1 does not alter translation rates, affect accumulation of ribosomal subunits, change mRNA poly(A) tail lengths, or result in a defect in mRNA decay. Rather, Pbp1p appears to function in the nucleus to promote proper polyadenylation. In the absence of Pbp1p, 3′ termini of pre-mRNAs are properly cleaved but lack full-length poly(A) tails. These effects suggest that Pbp1p may act to repress the ability of Pab1p to negatively regulate polyadenylation.

The Rox1 repressor of the Saccharomyces cerevisiae hypoxic genes is a specific DNA-binding protein with a high-mobility-group motif

1993 ◽  
Vol 13 (10) ◽  
pp. 6071-6078
Author(s):  
B Balasubramanian ◽  
C V Lowry ◽  
R S Zitomer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Binding ◽  
Binding Protein ◽  
Consensus Sequence ◽  
High Mobility ◽  
High Mobility Group ◽  
Upstream Region ◽  
Yeast Saccharomyces Cerevisiae ◽  
Amino Terminal

The ROX1 gene encodes a repressor of the hypoxic functions of the yeast Saccharomyces cerevisiae. The DNA sequence of the gene was determined and found to encode a protein of 368 amino acids. The amino-terminal third of the protein contains a high-mobility-group motif characteristic of DNA-binding proteins. To determine whether the Rox1 repressor bound DNA, the gene was expressed in Escherichia coli cells as a fusion to the maltose-binding protein and this fusion was partially purified by amylose affinity chromatography. By using a gel retardation assay, both the fusion protein and Rox1 itself were found to bind specifically to a synthetic 32-bp DNA containing the hypoxic consensus sequence. We assessed the role of the general repressor Ssn6 in ANB1 repression. An ANB1-lacZ fusion was expressed constitutively in an ssn6 deletion strain, and deletion of the Rox1 binding sites in the ANB1 upstream region did not increase the level of derepression, suggesting that Ssn6 exerts its effect through Rox1. Finally, ROX1 was mapped to yeast chromosome XVI, near the ARO7-OSM2 locus.

scholarly journals The Rox1 repressor of the Saccharomyces cerevisiae hypoxic genes is a specific DNA-binding protein with a high-mobility-group motif.

1993 ◽  
Vol 13 (10) ◽  
pp. 6071-6078 ◽  
Author(s):  
B Balasubramanian ◽  
C V Lowry ◽  
R S Zitomer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Binding ◽  
Binding Protein ◽  
Consensus Sequence ◽  
High Mobility ◽  
High Mobility Group ◽  
Upstream Region ◽  
Yeast Saccharomyces Cerevisiae ◽  
Amino Terminal

The ROX1 gene encodes a repressor of the hypoxic functions of the yeast Saccharomyces cerevisiae. The DNA sequence of the gene was determined and found to encode a protein of 368 amino acids. The amino-terminal third of the protein contains a high-mobility-group motif characteristic of DNA-binding proteins. To determine whether the Rox1 repressor bound DNA, the gene was expressed in Escherichia coli cells as a fusion to the maltose-binding protein and this fusion was partially purified by amylose affinity chromatography. By using a gel retardation assay, both the fusion protein and Rox1 itself were found to bind specifically to a synthetic 32-bp DNA containing the hypoxic consensus sequence. We assessed the role of the general repressor Ssn6 in ANB1 repression. An ANB1-lacZ fusion was expressed constitutively in an ssn6 deletion strain, and deletion of the Rox1 binding sites in the ANB1 upstream region did not increase the level of derepression, suggesting that Ssn6 exerts its effect through Rox1. Finally, ROX1 was mapped to yeast chromosome XVI, near the ARO7-OSM2 locus.

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