Interaction of formin FH2 with skeletal muscle actin. EPR and DSC studies.

Formins are highly conserved proteins that are essential in the formation and regulation of the actin cytoskeleton. The formin homology 2 (FH2) domain is responsible for actin binding and acts as an important nucleating factor in eukaryotic cells. In this work EPR and DSC were used to investigate the properties of the mDia1-FH2 formin fragment and its interaction with actin.

Adaptation to tert-butyl hydroperoxide at a plasma membrane level in the fission yeast Schizosaccharomyces pombe parental strain and its t-BuOOH-resistant mutant.

The one-gene mutant hyd1-190 of the fission yeast Schizosaccharomyces pombe displayed four-fold resistance to tert-butyl hydroperoxide (t-BuOOH) in comparison with its parental strain hyd(+). The cells of hyd1-190 exhibited a quantitative alteration in the sterol content and hence in the fatty acid composition of the plasma membrane, reflected in a two-fold amphotericin B sensitivity, increased rigidity of the plasma membrane, revealed by an elevated (Δ7.9 °C) phase-transition temperature, measured by means of electron paramagnetic resonance spectroscopy, and a significantly decreased uptake of glycerol.

Regulation of patulin-induced oxidative stress processes in the fission yeast Schizosaccharomyces pombe.

Patulin (PAT), is one of the most widely disseminated mycotoxins found in agricultural products. In this study the PAT-induced accumulation of reactive oxygen species (ROS) and the regulation of the specific activities of antioxidant enzymes were investigated in the single cell eukaryotic organism Schizosaccharomyces pombe. In comparison with the untreated cells, 500 μM PAT treatment caused a 43% decrease in the concentration of the main intracellular antioxidant, glutathione (GSH); this depletion of GSH initiated a 2.

In vivo direct patulin-induced fluidization of the plasma membrane of fission yeast Schizosaccharomyces pombe.

Patulin is a toxic metabolite produced by various species of Penicillium, Aspergillus and Byssochlamys. In the present study, its effects on the plasma membrane of fission yeast Schizosaccharomyces pombe were investigated. The phase-transition temperature (G) of untreated cells, measured by electron paramagnetic resonance spectrometry proved to be 14.

Oxidative stress induced by HIV-1 F34IVpr in Schizosaccharomyces pombe is one of its multiple functions.

Human immunodeficiency virus type 1 (HIV-1) viral protein R (Vpr) exerts multiple effects on viral and host cellular activities during infection, including induction of cell cycle G(2) arrest and cell death in both human and the fission yeast Schizosaccharomyces pombe cells. In this study, a mutant derivative of Vpr (F34IVpr), which causes transient G2 arrest with little or no effect of cell killing, was used to study the molecular impact of Vpr on cellular oxidative stress responses in S. pombe.

The uncoupling of the effects of formins on the local and global dynamics of actin filaments.

In this study, experiments were carried out in the conventional and saturation-transfer electron paramagnetic resonance (EPR) time domains to explore the effect of mDia1-FH2 formin fragments on the dynamic and conformational properties of actin filaments. Conventional EPR measurements showed that addition of formin to actin filaments produced local conformational changes in the vicinity of Cys-374 by increasing the flexibility of the protein matrix in the environment of the label. The results indicated that it was the binding of formin to the barbed end that resulted in these conformational changes.

High-dose methylprednisolone influences the physiology and virulence of Candida albicans ambiguously and enhances the candidacidal activity of the polyene antibiotic amphotericin B and the superoxide-generating agent menadione.

Although exposure of Candida albicans cells to high-dose (4 mM) methylprednisolone stimulated microbial growth, germination rate in serum and phospholipase release, it also promoted the recognition of C. albicans cells by polymorphonuclear leukocytes. Pretreatment of C.

Intermonomer cross-linking of F-actin alters the dynamics of its interaction with H-meromyosin in the weak-binding state.

Previous cross-linking studies [Kim E, Bobkova E, Hegyi G, Muhlrad A & Reisler E (2002) Biochemistry 41, 86-93] have shown that site-specific cross-linking among F-actin monomers inhibits the motion and force generation of actomyosin. However, it does not change the steady-state ATPase parameters of actomyosin. These apparently contradictory findings have been attributed to the uncoupling of force generation from other processes of actomyosin interaction as a consequence of reduced flexibility at the interface between actin subdomains-1 and -2.

Involvement of serum retinoids and Leiden mutation in patients with esophageal, gastric, liver, pancreatic, and colorectal cancers in Hungary.

Aim: To analyze the serum levels of retinoids and Leiden mutation in patients with esophageal, gastric, liver, pancreatic, and colorectal cancers.

Methods: The changes in serum levels of retinoids (vitamin A, alpha- and beta-carotene, alpha- and beta-cryptoxanthin, zeaxanthin, lutein) and Leiden mutation were measured by high liquid performance chromatography (HPLC) and polymerase chain reaction (PCR) in 107 patients (70 males/37 females) with esophageal (0/8), gastric (16/5), liver (8/7), pancreatic (6/4), and colorectal (30/21 including 9 patients suffering from in situ colon cancer) cancer. Fifty-seven healthy subjects (in matched groups) for controls of serum retinoids and 600 healthy blood donors for Leiden mutation were used.

Scavenger capacity of follicular fluid, decidua and culture medium with regard to assisted reproduction: an in vitro study using electron paramagnetic resonance spectroscopy.

The natural scavenger capacity of follicular fluid of women pre-treated for in vitro fertilization (IVF) embryo transfer, Ham's F10 nutrient mixture used for oocyte culture and endometrium samples were studied in a hydroxyl free radical generating system, using electron paramagnetic resonance (EPR) spectroscopy. The EPR signals appearing after the addition of samples to the reaction mixture show a diversely decreasing production of phenyl-tertier-butylnitrone spin adducts. Presumably, the follicular fluid and endometrium samples contain active factors which function as radical scavengers.

Chromate tolerance caused by reduced hydroxyl radical production and decreased glutathione reductase activity in Schizosaccharomyces pombe.

The stable Cr(VI)-tolerant chr1-66T mutant of Schizosaccharomyces pombe, which carries one simple gene mutation responsible for Cr(VI) tolerance, accumulated and reduced the chromate anion (CrO(4)(2-)) significantly more slowly than did its parental strain 6chr(+). The mutant chr1-66T proved to be sensitive to oxidative stressors such as H(2)O(2), menadione, tert-butyl hydroperoxide and Cd(2+). Both the Cr(VI) tolerance and the oxidative stress sensitivity were attributed to a decreased specific glutathione reductase activity.

Chromate sensitivity in fission yeast is caused by increased glutathione reductase activity and peroxide overproduction.

The Cr(VI)-sensitive mutant chr-51S of the Schizosaccharomyces pombe accumulated chromate (CrO(4) (2-)) and reduced Cr(V) to much greater extent, than did its parental strain 6 chr(+). Sublethal doses of K(2)Cr(2)O(7) did not induce any adaptive stress response, while H(2)O(2) or menadione pretreatment proved protective against the cell injuries caused by Cr(VI). The intracellular GSH concentration in chr-51S cells was approximately half of that for the 6 chr(+).