We have constructed a series of plasmids containing modified form the phoA gene Escherichia coli K-12 that general utility for studies protein secretion. In these plasmids, promoter and signal sequence-encoding region been deleted; thus, expression gene, giving rise to active alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1], is absolutely dependent upon fusion in correct reading frame DNA promoter, translational start site, complete region....

SUMMARY The fungistatic properties of the normal saturated fatty acids containing from one to 14 carbon atoms have been studied over a pH range 2 8. Many these exhibit remarkable effectiveness in inhibition mold growth. This varies according chain length, concentration acid, and media. A branched general, is less effective fungistatically than corresponding straight acid. Unsaturation tends increase an At neutrality, eight 12 are most for biochemical fact suggested that nutritional value be...

Transcription of the fbp1 gene, encoding fructose-1,6-bisphosphatase, Schizosaccharomyces pombe is subject to glucose repression. Previous work has demonstrated that several genes (git genes) are required for this In report we demonstrate one these genes, git2, same as cyr1 which encodes adenylate cyclase, and loss-of-function mutations in git2 cause constitutive transcription. Addition cAMP growth medium suppresses transcriptional defect mutants well strains carry any six additional git...

Abstract The fission yeast Schizosaccharomyces pombe responds to environmental glucose by activating adenylate cyclase. resulting cAMP signal activates protein kinase A (PKA). PKA inhibits starvation-induced processes, such as conjugation and meiosis, the transcription of fbp1 gene that encodes gluconeogenic enzyme fructose-1,6-bisphosphatase. We previously identified a collection git genes required for repression transcription, including pka1/git6, encoding catalytic subunit, git2/cyr1,...

Glucose is the preferred carbon and energy source in prokaryotes, unicellular eukaryotes, metazoans. However, excess of glucose has been associated with several diseases, including diabetes less understood process aging. On contrary, limiting (i.e., calorie restriction) slows aging age-related diseases most species. Understanding mechanism by which limits life span therefore important for any attempt to control diseases. Here, we use yeast Schizosaccharomyces pombe as a model study...

Molecular studies in yeast often require the isolation of both plasmid and chromosomal DNA. Plasmid DNA is used transformation E. coli, whereas for Southern hybridization analysis, vitro amplification by polymerase chain reaction (PCR), or cloning integrated plasmids. This unit presents two variations "smash grab" protocol that produce suitable all these applications. These protocols work Saccharomyces cerevisiae Schizosaccharomyces pombe.

Transcription of the fbp1 gene Schizosaccharomyces pombe, encoding fructose-1,6-bisphosphatase, is glucose repressible. We have constructed two hybrid genes, containing promoter, that allow selection for mutations alter transcriptional regulation fbp1. Strains carrying fbp1-ura4 and fbp1-lacZ fusions are phenotypically Ura-, resistant to 5-fluoro-orotic acid, express a low level beta-galactosidase activity when grown under repressing conditions (8% glucose). By selecting Ura+ strains...

Abstract Since its humble start as a model organism in two European laboratories the 1940s and 1950s, fission yeast Schizosaccharomyces pombe has grown to become one of best-studied eukaryotes today. This article outlines way which interest S. developed spread from Europe Japan, North America, elsewhere beginnings up first International Meeting devoted this 1999. We describe expansion research during period with an emphasis on many individual researchers involved their interactions that...

An important eukaryotic signal transduction pathway involves the regulation of effector enzyme adenylate cyclase, which produces second messenger, cAMP. Previous genetic analyses demonstrated that glucose repression transcription Schizosaccharomyces pombe fbp1 gene requires function encoded by git2 gene. As mutations in and six additional git genes are suppressed exogenous cAMP, these 'upstream' were proposed to act produce a glucose-induced cAMP signal. We report here assays levels...

Schizosaccharomyces pombe regulates intracellular cAMP levels, and thus cAMP-dependent protein kinase (PKA) activity, in response to changes nutrient conditions. Mutations any of eight git genes inhibit glucose repression fbp1 transcription, alter the cell morphology, cause a reduction growth rate. The encode components an adenylate cyclase activation pathway, itself, catalytic subunit PKA. Three these have been identified other studies as regulators meiosis. Here we show that sck1 gene,...

A significant challenge to our understanding of eukaryotic transcriptional regulation is determine how multiple signal transduction pathways converge on a single promoter regulate transcription in divergent fashions. To study this, we have investigated the Schizosaccharomyces pombe fbp1 gene that repressed by cyclic AMP (cAMP)-dependent protein kinase (PKA) pathway and activated stress-activated mitogen-activated (MAPK) pathway. In this study, identified characterized twocis-acting elements...

The Hat1 histone acetyltransferase catalyzes the acetylation of H4 at lysines 5 and 12, same sites that are acetylated in newly synthesized H4. By performing (HAT) assays on various synthetic N-terminal peptides, we have examined interactions between tail domain. It was found requires presence positively charged amino acids positions 8 16 H4, normally occupied by lysine; however, lysine per se is not essential can be replaced arginine. In contrast, replacing Lys-8 -16 with glutamines reduces...

It has been postulated that a myriad of long noncoding RNAs (lncRNAs) contribute to gene regulation.In fission yeast, glucose starvation triggers lncRNA transcription across promoter regions stress-responsive genes including fbp1 (fructose-1,6-bisphosphatase1).At the promoter, this promotes chromatin remodeling and mRNA expression.Here, we demonstrate such upstream facilitates association transcriptional activator Atf1 at sites transcription, leading activation downstream stress...

Abstract The Schizosaccharomyces pombe fbp1 gene, which encodes fructose-1,6-bis-phosphatase, is transcriptionally repressed by glucose through the activation of cAMP-dependent protein kinase A (PKA) and activated starvation a mitogen-activated (MAPK). To identify transcriptional regulators acting downstream from or in parallel to PKA, we screened an adh-driven cDNA plasmid library for genes that increase transcription strain with elevated PKA activity. Two such clones express...

In the fission yeast Schizosaccharomyces pombe, genetic studies have identified genes that are required for glucose repression of fbp1 transcription. The git2 gene, also known as cyr1, encodes adenylate cyclase. Adenylate cyclase converts ATP into second messenger cAMP part many eukaryotic signal transduction pathways. git1, git3, git5, git7, git8 and git10 act upstream cyclase, presumably encoding an activation pathway. mammalian cells, enzymatic activity is regulated by heterotrimeric...

Abstract Fission yeast adenylate cyclase, like mammalian cyclases, is regulated by a heterotrimeric G protein. The gpa2 Gα and git5 Gβ are both required for glucose-triggered cAMP signaling. unique member of the family in that it lacks an amino-terminal coiled-coil domain shown to be essential folding interaction with Gγ subunits. Using bait two-hybrid screen, we identified git11 gene. Co-immunoprecipitation studies confirm composition this Gβγ dimer. Cells deleted defective glucose...