S phase and mitotic onset are brought about by the action of multiple different cyclin-CDK complexes. However, it has been suggested that changes in total level CDK kinase activity, rather than substrate specificity, drive temporal ordering mitosis. Here, we present a phosphoproteomics-based systems analysis substrates fission yeast demonstrate phosphorylation can be temporally ordered during cell cycle single cyclin-CDK. This is achieved rising activity differential sensitivity to over wide...

Significance Although gamma delta (γδ) T cells compose an evolutionarily conserved third lineage of diversified lymphocytes, alongside αβ and B cells, they can seem overtly different across species tissues. Thus, human blood γδ show butyrophilin (BTN)3A1-dependent responses to metabolites (“phosphoantigens”) not seen by rodent whereas some rodent, γδ-rich compartments, notably in the skin, lack obvious counterparts. Recently, however, mouse intraepithelial gut were found be regulated...

Highlights•Global analysis of phosphorylation dynamics during the fission yeast cell cycle•Reveals kinase-specific waves throughout interphase and mitosis•Mitotic kinases show significantly different dependencies on upstream CDK activity•Kinases directly downstream mediate earlier mitotic phosphorylationSummaryMultiple protein regulate cell-cycle progression, which cyclin-dependent (CDKs) are thought to act as master regulators. We have used quantitative phosphoproteomics analyze cycle at...

Abstract Cyclin-dependent kinases (CDKs) lie at the heart of eukaryotic cell cycle control, with different cyclin–CDK complexes initiating DNA replication (S-CDKs) and mitosis (M-CDKs) 1,2 . However, principles on which organize temporal order events are contentious 3 One model proposes that S-CDKs M-CDKs functionally specialized, substantially substrate specificities to execute 4–6 A second redundant each other, both acting as sources overall CDK activity 7,8 In this model, increasing...

How cells control the overall size and growth of membrane-bound organelles is an important unanswered question cell biology. Fission yeast maintain a nuclear proportional to cellular size, resulting in constant ratio between volumes (N/C ratio). We have conducted genome-wide visual screen fission gene deletion collection for viable mutants altered their N/C ratio, found that defects both nucleocytoplasmic mRNA transport lipid synthesis alter ratio. Perturbing export results accumulation...

Cyclic nucleotide signalling is a major regulator of malaria parasite differentiation. Phosphodiesterase (PDE) enzymes are known to control cyclic GMP (cGMP) levels in the parasite, but mechanisms by which AMP (cAMP) regulated remain enigmatic. Here, we demonstrate that Plasmodium falciparum phosphodiesterase β (PDEβ) hydrolyses both cAMP and cGMP essential for blood stage viability. Conditional gene disruption causes profound reduction invasion erythrocytes rapid death those merozoites...

A new peptide sequence (MB1) has been designed which, in the presence of a trivalent lanthanide ion, programmed to self-assemble form three stranded metallo-coiled coil, Ln(III)(MB1)3. The binding site incorporated into hydrophobic core using natural amino acids, restricting water access lanthanide. resulting terbium coiled coil displays luminescent properties consistent with lack first coordination sphere molecules. Despite this gadolinium be reported, promising magnetic resonance contrast...

Abstract The functions of cohesin are central to genome integrity, chromosome organization and transcription regulation through its prevention premature sister-chromatid separation the formation DNA loops. loading onto chromatin depends on Scc2–Scc4 complex; however, little is known about how it stimulates cohesion-loading activity. Here we determine large ‘hook’ structure Scc2 responsible for catalysing loading. We identify key surfaces that crucial in vivo . With aid previously determined...

Resource12 April 2018Open Access Source DataTransparent process Phosphoproteome dynamics during mitotic exit in budding yeast Sandra A Touati Chromosome Segregation Laboratory, The Francis Crick Institute, London, UK Search for more papers by this author Meghna Kataria Andrew W Jones Mass Spectrometry Proteomics Science Technology Platform, Ambrosius P Snijders Frank Uhlmann Corresponding Author [email protected] orcid.org/0000-0002-3527-6619 protected]crick.ac.uk Information Touati1,...

Temporal control over protein phosphorylation and dephosphorylation is crucial for accurate chromosome segregation completion of the cell division cycle during exit from mitosis. In budding yeast, Cdc14 phosphatase thought to be a major regulator at this time, while in higher eukaryotes PP2A phosphatases take dominant role. Here, we use time-resolved phosphoproteome analysis yeast evaluate respective contributions Cdc14, PP2ACdc55, PP2ARts1. This reveals an overlapping requirement all three...

In vivo protein nitration is associated with many disease conditions that involve oxidative stress and inflammatory response. The modification involves addition of a nitro group at the position ortho to phenol tyrosine give 3-nitrotyrosine. To understand mechanisms consequences nitration, it necessary develop methods for identification nitrotyrosine-containing proteins localization sites modification. Here, we have investigated electron capture dissociation (ECD) collision-induced (CID)...

Highlights•The hydrophobic patch of human and yeast cyclin B directs it to the centrosome•Loss allows S phase but prevents mitosis•Compartmentalized mitotic CDK phosphorylation relies on patchSummaryThe cyclin-dependent kinases (CDKs) are major cell-cycle regulators that phosphorylate hundreds substrates, controlling onset M [1–3]. However, patterns substrate increase not uniform, as different substrates become phosphorylated at times cells proceed through cell cycle [4, 5]. In fission...

The PKCε-regulated genome protective pathway provides transformed cells a failsafe to successfully complete mitosis. Despite the necessary role for Aurora B in this programme, it is unclear whether its requirement sufficient or if other PKCε cell cycle targets are involved. To address this, we developed trapping strategy using UV-photocrosslinkable amino acids encoded kinase domain. validation of mRNA binding protein SERBP1 as substrate revealed series mitotic events controlled by catalytic...

Abstract A fundamental feature of cellular growth is that protein and RNA amounts scale with cell size so concentrations remain constant. key component to this global transcription rates increase in larger cells, but the underlying mechanism has remained unknown. Here, we identify RNAPII as major limiting factor increasing budding yeast highly sensitive dosage not other components general transcriptional machinery. Our experiments support a dynamic equilibrium model where at given set by...

Abstract The cohesin complex participates in many structural and functional aspects of genome organization. Cohesin recruitment onto chromosomes requires nucleosome-free DNA the Scc2-Scc4 loader that catalyzes topological loading. Additionally, facilitates promoter nucleosome clearance a yet unknown way, it recognizes chromatin receptors such as RSC remodeler. Here, we explore loader-RSC interaction. Amongst multi-pronged contacts by Scc2 Scc4, find Scc4 conserved patch on ATPase motor...

Electroconvulsive therapy (ECT) is one of the most controversial treatments in psychiatry. This controversy and diverse often strongly held opinions can make decision making processes around ECT more complex. consumer-led project explored experiences individuals who had received terms information they received, their experience suggestions for ways that be improved. Interviews were conducted by consumer researchers also transcripts analysed using constant comparative techniques. Seventeen...

Abstract Aim Inflammatory periodontal disease is widespread in dogs. This study evaluated site‐specific changes the canine gingival crevicular fluid ( GCF ) proteome during longitudinal progression from very mild gingivitis to periodontitis. Periodontitis diagnosis dogs requires general anaesthesia with associated risks and costs; our ultimate aim was develop a periodontitis diagnostic for application conscious The objective of this work identify potential biomarkers Material Methods...

Cell growth is orchestrated by a number of interlinking cellular processes. Components the TOR pathway have been proposed as potential regulators cell growth, but little known about their immediate effects on protein synthesis in response to TOR-dependent inhibition. Here, we present resource providing an in-depth characterisation Schizosaccharomyces pombe phosphoproteome relation changes observed global upon We find that after inhibition, rate rapidly reduced and notable phosphorylation are...

Abstract Sister-chromatid cohesion is established by Eco1-mediated acetylation on two conserved tandem lysines in the cohesin Smc3 subunit. However, molecular basis of Eco1 substrate recognition and not fully understood. Here, we discover rationalize specificity using mass spectrometry coupled with in-vitro assays crystallography. Our structures X. laevis Eco2 (xEco2) bound to its primary secondary substrates demonstrate plasticity substrate-binding site, which confers concerted...

Electron capture dissociation mass spectrometry offers several advantages for the analysis of peptides, most notably that backbone c and z fragments typically retain labile modifications such as phosphorylation. We have shown previously that, in some cases, presence phosphorylation has a deleterious effect on peptide sequence coverage, hypothesized intramolecular interactions involving phosphate group were preventing separation fragments. In present work, we seek to rationalize observed ECD...

Previously we have shown that the presence of 3-nitrotyrosine within a peptide sequence severely depletes backbone fragments typically observed following electron capture dissociation (ECD) mass spectrometry. Instead, ECD nitrated peptides is characterised by abundant losses small neutrals (hydroxyl radicals, water and ammonia). Here, investigate origin ammonia loss comparing behaviour lysine- arginine-containing peptides, their N-acetylated counterparts, containing no basic amino acid...