Fission yeast 26S proteasome mutants are multi-drug resistant due to stabilization of the pap1 transcription factor

Here we report the result of a genetic screen for mutants resistant to the microtubule poison methyl benzimidazol-2-yl carbamate (MBC) that were also temperature sensitive for growth. In total the isolated mutants were distributed in ten complementation groups. Cloning experiments revealed that most of the mutants were in essential genes encoding various 26S proteasome subunits. We found that the proteasome mutants are multi-drug resistant due to stabilization of the stress-activated transcription factor Pap1. We show that the ubiquitylation and ultimately the degradation of Pap1 depend on the Rhp6/Ubc2 E2 ubiquitin conjugating enzyme and the Ubr1 E3 ubiquitin-protein ligase. Accordingly, mutants lacking Rhp6 or Ubr1 display drug-resistant phenotypes

A co-development approach to conservation leads to informed habitat design and rapid establishment of amphibian communities

1. Effective wildlife restoration is a critical requirement of many conservation actions. The outcome of conservation interventions can be optimised through knowledge of species’ habitat requirements, but few studies consider the impact of using explicit evidence from dedicated local research to inform the design phase of habitat management. Furthermore, interventions administered externally from the top down, whilst simpler than those developed in discussion with multiple stakeholders including land managers (i.e. co-development), run the risk of failing to engage local people. 2. In this study, we focus on interventions in the Scottish Highlands to improve the availability and suitability of breeding ponds for local amphibian assemblages. We collected and analysed data based on 129 ecological variables across 88 reference ponds to quantify the local habitat preferences. We used the findings from these analyses to inform the construction or restoration of 25 intervention ponds co-developed in partnership with stakeholders (landowners, foresters, citizen scientists and government agencies). Following the interventions, we monitored amphibian communities at these sites over 4 years. We assessed presence and abundance of all five native amphibians (the anurans Rana temporaria and Bufo bufo, and the salamanders Lissotriton helveticus, L. vulgaris and Triturus cristatus) using egg searching, dip-netting, torching and trapping. 3. The new habitats were overall characterised by ecological conditions more favourable to amphibians than the reference ponds. We recorded a total of 51 colonisation events. Within two breeding seasons after construction or restoration, the intervention ponds hosted the full complement of species, mirroring amphibian diversity patterns found in the local reference ponds. 4. Our study shows that ecological research to quantify local habitat requirements and working with commercial land-managers to ensure equitable benefits prior to designing conservation actions can promote rapid and efficient recovery of wildlife

The Fission Yeast Homeodomain Protein Yox1p Binds to MBF and Confines MBF-Dependent Cell-Cycle Transcription to G1-S via Negative Feedback

The regulation of the G1- to S-phase transition is critical for cell-cycle progression. This transition is driven by a transient transcriptional wave regulated by transcription factor complexes termed MBF/SBF in yeast and E2F-DP in mammals. Here we apply genomic, genetic, and biochemical approaches to show that the Yox1p homeodomain protein of fission yeast plays a critical role in confining MBF-dependent transcription to the G1/S transition of the cell cycle. The yox1 gene is an MBF target, and Yox1p accumulates and preferentially binds to MBF-regulated promoters, via the MBF components Res2p and Nrm1p, when they are transcriptionally repressed during the cell cycle. Deletion of yox1 results in constitutively high transcription of MBF target genes and loss of their cell cycle–regulated expression, similar to deletion of nrm1. Genome-wide location analyses of Yox1p and the MBF component Cdc10p reveal dozens of genes whose promoters are bound by both factors, including their own genes and histone genes. In addition, Cdc10p shows promiscuous binding to other sites, most notably close to replication origins. This study establishes Yox1p as a new regulatory MBF component in fission yeast, which is transcriptionally induced by MBF and in turn inhibits MBF-dependent transcription. Yox1p may function together with Nrm1p to confine MBF-dependent transcription to the G1/S transition of the cell cycle via negative feedback. Compared to the orthologous budding yeast Yox1p, which indirectly functions in a negative feedback loop for cell-cycle transcription, similarities but also notable differences in the wiring of the regulatory circuits are evident

Comparative genetic, proteomic and phosphoproteomic analysis of C. <i>elegans </i>embryos with a focus on <i>ham</i>-1/STOX and <i>pig</i>-1/MELK in dopaminergic neuron development

Asymmetric cell divisions are required for cellular diversity and defects can lead to altered daughter cell fates and numbers. In a genetic screen for C. elegans mutants with defects in dopaminergic head neuron specification or differentiation, we isolated a new allele of the transcription factor HAM-1 [HSN (Hermaphrodite-Specific Neurons) Abnormal Migration]. Loss of both HAM-1 and its target, the kinase PIG-1 [PAR-1(I)-like Gene], leads to abnormal dopaminergic head neuron numbers. We identified discrete genetic relationships between ham-1, pig-1 and apoptosis pathway genes in dopaminergic head neurons. We used an unbiased, quantitative mass spectrometry-based proteomics approach to characterise direct and indirect protein targets and pathways that mediate the effects of PIG-1 kinase loss in C. elegans embryos. Proteins showing changes in either abundance, or phosphorylation levels, between wild-type and pig-1 mutant embryos are predominantly connected with processes including cell cycle, asymmetric cell division, apoptosis and actomyosin-regulation. Several of these proteins play important roles in C. elegans development. Our data provide an in-depth characterisation of the C. elegans wild-type embryo proteome and phosphoproteome and can be explored via the Encyclopedia of Proteome Dynamics (EPD) - an open access, searchable online database

Origin of Irreversibility of Cell Cycle Start in Budding Yeast

In budding yeast, the commitment to entry into a new cell division cycle is made irreversible by positive feedback-driven expression of the G1 cyclins Cln1,2

Networking Our Way to Better Ecosystem Service Provision.

The ecosystem services (EcoS) concept is being used increasingly to attach values to natural systems and the multiple benefits they provide to human societies. Ecosystem processes or functions only become EcoS if they are shown to have social and/or economic value. This should assure an explicit connection between the natural and social sciences, but EcoS approaches have been criticized for retaining little natural science. Preserving the natural, ecological science context within EcoS research is challenging because the multiple disciplines involved have very different traditions and vocabularies (common-language challenge) and span many organizational levels and temporal and spatial scales (scale challenge) that define the relevant interacting entities (interaction challenge). We propose a network-based approach to transcend these discipline challenges and place the natural science context at the heart of EcoS research.The QUINTESSENCE Consortium gratefully acknowledges the support of Départment SPE and Métaprogramme ECOSERV of INRA, and the French ANR projects PEERLESS (ANR-12-AGRO-0006) and AgroBioSE (ANR-13-AGRO-0001).This is the final version of the article. It first appeared from Elsevier via http://dx.doi.org/10.1016/j.tree.2015.12.00

Daughter-Specific Transcription Factors Regulate Cell Size Control in Budding Yeast

The asymmetric localization of cell fate determinants results in asymmetric cell cycle control in budding yeast