Septins constitute a group of GTP-binding proteins involved in cytokinesis and other essential cellular functions. They form heterooligomeric complexes that polymerize into nonpolar filaments are dynamic during different stages the cell cycle. Posttranslational modifications interacting partners widely accepted regulators septin filament function, but contribution nucleotide is undefined due to lack detailed structural information. Previous low-resolution structures showed G domain assembles...

Septins are guanine nucleotide-binding proteins that polymerize into filamentous and higher-order structures. Cdc42 its effector Gic1 involved in septin recruitment, ring formation dissociation. The regulatory mechanisms behind these processes not well understood. Here, we have used electron microscopy cryo tomography to elucidate the structural basis of Gic1-septin Gic1-Cdc42-septin interaction. We show acts as a scaffolding protein for filaments forming long flexible filament cables....

The NUM1 gene is involved in the control of nuclear migration Saccharomyces cerevisiae. content mRNA fluctuates during cell cycle, reaching a maximum at S/G2 phase, and translation product Num1p associates with cortex mother cells mainly S, G2, mitosis, as seen by indirect immunofluorescence. spindle NUM1-deficient large-budded often fails to align along mother/bud axis, while abnormally elongated astral microtubules emanate from both pole bodies. A num1 null mutation confers temperature...

Num1p, a cortical 313-kD protein, controls cytoplasmic microtubule (cMT) functions and nuclear migration through the bud neck in anaphase cells. A green fluorescent protein (GFP)-Num1p fusion localizes at tip distal mother pole of living cells, apparently forming cMT capture sites late anaphase. In addition, galactose-induced GFP-Num1p is seen lateral regions cortex. The location Num1p depends on Bni1p but does not require Kar9p, Dyn1p, or cMTs, whereas contacts with polar dots are reduced...

Septins are filament-forming GTPases involved in cytokinesis and cortical organization. In the yeast Saccharomyces cerevisiae, septins encoded by CDC3, CDC10, CDC11, CDC12 form a high-molecular-weight complex, localized at cytoplasmic face of plasma membrane mother-bud neck. While septin function cellular level is fairly well understood, progress on structure-function analysis these proteins has been slow limited lack large amounts pure complex. monomeric apparently non-native aggregates,...

The hormone receptor-like protein Gpr1p physically interacts with phosphatidylinositol-specific phospholipase C (Plc1p) and the Gα Gpa2p, as shown by two-hybrid assays co-immune precipitation of epitope-tagged proteins. Plc1p binds to in either presence or absence Gpa2, whereas Gpr1p/Gpa2p association depends on Plc1p. Genetic interactions between null mutations plc1Δ,gpr1Δ, gpa2Δ, ras2Δsuggest that acts together Gpa2p a growth control pathway operating parallel Ras2p function. Diploid cells...

RanBP type proteins have been reported to increase the catalytic efficiency of RanGAP-mediated GTPase reaction on Ran. Since structure Ran-RanBP1-RanGAP complex showed RanBP1 be located away from active site, we reinvestigated using fluorescence spectroscopy under pre-steady-state conditions. We can show that indeed does not influence rate-limiting step reaction, which is cleavage GTP and/or release product Pi. It does, however, dynamics Ran-RanGAP interaction, its most dramatic effect being...

Live-imaging analysis is performed in many laboratories all over the world. Various tools have been developed to enable protein labeling either plasmid or genomic context live yeast cells. Here, we introduce a set of nine integrative modules for C-terminal gene tagging that combines three fluorescent proteins (FPs)—ymTagBFP, mCherry and yTagRFP-T with dominant selection markers: geneticin, nourseothricin hygromycin. In addition, construction two episomal Saccharomyces cerevisiae photostable...

Abstract Septins are a family of conserved cytoskeletal proteins playing an essential role in cytokinesis and many other cellular processes fungi animals. In budding yeast Saccharomyces cerevisiae , septins form filaments higher-order structures at the mother-bud neck depending on particular stage cell cycle. Septin division plane serve as scaffold to recruit required for processes. The formation localization septin stages cycle also determine functionality these proteins. Many different...

Mining of natural sources for new secondary metabolites has a successful history, which is reflected by the fact that over 50% all drugs, currently on market, are derived from products. Bacteria one most important bioactive products destined drug discovery. However, less than 1% microorganisms observed in different habitats have been cultivated and characterized. To explore genomic functional diversity vast majority microbial world, novel methods were introduced, based analysis DNA isolated...

We previously identified a polyketide synthase gene cluster, aur1, responsible for the production of angucycline antibiotic auricin in Streptomyces aureofaciens CCM 3239. A sequence analysis aur1 flanking regions revealed presence several genes encoding proteins homologous to those linear plasmid replication, partitioning and telomere-binding. Pulse-field gel electrophoresis detected single, 240-kb plasmid, pSA3239, S. CCM3239. The cluster pSA3239 was confirmed by approaches. In addition...

SUMMARY: Conditions for the preparation of protoplasts from Streptomyces aureofaciens and transformation with several replicative integrative plasmids were optimized. Of all used, carrying various replication origins, only plasmid pGM9 produced transformants, a efficiency 2 × 103 (μg DNA)-1. When DNA isolated S. was increased to 105 transformants Transformation containing part hrdA gene gave rise an approximately 30 Integration frequency strongly reduced when modified by dam or dcm...

Bifunctional catalase-peroxidases (KatGs) are heme oxidoreductases widely spread among bacteria, archaea and lower eukaryotes. In fungi, two KatG groups with different localization have evolved, intracellular (KatG1) extracellular (KatG2) proteins. Here, the cloning, expression analysis subcellular of novel katG1 genes from soil fungi Chaetomium globosum cochliodes reported. Whereas, metalloenzyme Ch. is expressed constitutively, KatG1 reveals a slight increase in after induction oxidative...

Using antibody-linked polymerase assay we studied the polypeptide composition of DNA-dependent RNA from Streptomyces aureofaciens and immunological cross-reaction with Escherichia coli polymerase. We identified about 25 'ALPA-reactive' polypeptides which are probably involved in transcriptional apparatus. demonstrated that beta' beta subunits S. E. immunologically related sigma70 (E. coli) shows immunochemical similarity sigma35 (S. aureofaciens). According to reconstitution holoenzyme sigma...