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Ran An, Li Cheng, Lijian Chen, Jian Du
Ring finger protein 2 (RNF2), also known as RING2 or RING1B, displays oncogenic functions in different types of cancers, yet, the function of RNF2 during mitosis has not been evaluated. A yeast two‑hybrid screen was undertaken using a human HeLa cDNA library to explore and identify proteins that interact with RNF2. Several positive clones, including Polo‑like kinase 1 (Plk1), a critical regulator of mitosis, were identified. The interaction between RNF2 and Plk1 was confirmed using a β‑galactosidase and growth test in selective media, in vitro glutathione S‑transferase pull‑down, and in vivo immunoprecipitation assays...
May 2018: Oncology Reports
Srinivasan Ramakrishnan, Beejan Asady, Roberto Docampo
Membrane contact sites are regions of close apposition between two organelles, typically less than 30 nanometers apart, that facilitate transfer of biomolecules. The presence of contact sites has been demonstrated in yeast, plants, and mammalian cells. Here, we investigated the presence of such contact sites in Trypanosoma brucei . In mammalian cells, endoplasmic reticulum-mitochondria contact sites facilitate mitochondrial uptake of Ca2+ released by the ER-located inositol 1,4,5-trisphosphate receptor (InsP₃R)...
March 22, 2018: Pathogens
Hongmin Hou, Hui Jia, Qin Yan, Xiping Wang
Salinity and drought are two major abiotic stresses that limit grape productivity. Responses to stress in grape are known to be regulated by several families of transcription factors. However, little is known about the role of grape Squamosa promoter binding protein (SBP)-box transcription factor genes in response to abiotic stress. To better understand the functions of the grape SBP-box genes in abiotic stress tolerance, a full-length complementary DNA (cDNA) sequence of the putative SBP-box transcription factor gene, VpSBP16 was amplified from Chinese wild grapevine Vitis pseudoreticulata clone "Baihe-35-1"...
March 22, 2018: International Journal of Molecular Sciences
Joana Rodrigues, David Lydall
Saccharomyces cerevisiae is a commonly used model organism for understanding eukaryotic gene function. However, the close proximity between yeast genes can complicate the interpretation of yeast genetic data, particularly high-throughput data. In this study, we examined the interplay between genes encoding components of the PAF1 complex and VPS36, the gene located next to CDC73 on chromosome XII. The PAF1 complex (Cdc73, Paf1, Ctr9, Leo1, and Rtf1, in yeast) affects RNA levels by affecting transcription, histone modifications, and post-transcriptional RNA processing...
March 22, 2018: Current Genetics
Siobhan A Turner, Qinxi Ma, Mihaela Ola, Kontxi Martinez de San Vicente, Geraldine Butler
Fungi can use a wide variety of nitrogen sources. In the absence of preferred sources such as ammonium, glutamate, and glutamine, secondary sources, including most other amino acids, are used. Expression of the nitrogen utilization pathways is very strongly controlled at the transcriptional level. Here, we investigated the regulation of nitrogen utilization in the pathogenic yeast Candida parapsilosis . We found that the functions of many regulators are conserved with respect to Saccharomyces cerevisiae and other fungi...
March 2018: MSphere
Philipp Wiemann, Alexandra A Soukup, Jacob S Folz, Pin-Mei Wang, Andreas Noack, Nancy P Keller
Background: Sequencing of fungal species has demonstrated the existence of thousands of putative secondary metabolite gene clusters, the majority of them harboring a unique set of genes thought to participate in production of distinct small molecules. Despite the ready identification of key enzymes and potential cluster genes by bioinformatics techniques in sequenced genomes, the expression and identification of fungal secondary metabolites in the native host is often hampered as the genes might not be expressed under laboratory conditions and the species might not be amenable to genetic manipulation...
2018: Fungal Biology and Biotechnology
Yoko Motokawa, Michifumi Kokubo, Nobuo Kuwabara, Ken-Ichiro Tatematsu, Hideki Sezutsu, Hideyuki Takahashi, Koichi Sakakura, Kazuaki Chikamatsu, Shigeki Takeda
Recent clinical trials with the aim of developing tumor antigen (TA)-specific cancer vaccines against a number of malignancies have focused on the identification of TAs presented by tumor cells and recognized by T cells. In the present study, the TA melanoma antigen family A4 (MAGE-A4) protein was produced using a transgenic (TG) silkworm system. Using in vitro stimulation, it was subsequently determined whether MAGE-A4 protein induced MAGE-A4-specific T cells from peripheral blood mononuclear cells of healthy donors...
March 2018: Experimental and Therapeutic Medicine
J M M Adams, A L Winters, E M Hodgson, J A Gallagher
Background: Energy crops including Miscanthus provide a storable, portable energy source which can be used to complement a wide range of products and energy generation systems. Miscanthus is predominantly used in Europe as a combustion material for electricity generation but also has the potential for biochemical conversion due to its high yield and low-nutrient requirements. The ratio of holocellulose (hemicellulose and cellulose combined) to acid detergent lignin (H:L) within the senesced material has previously been shown to indicate the relative suitability of Miscanthus accessions for thermochemical conversion...
2018: Biotechnology for Biofuels
Yingbo Liang, Shichun Cui, Xiaoli Tang, Yi Zhang, Dewen Qiu, Hongmei Zeng, Lihua Guo, Jingjing Yuan, Xiufen Yang
PevD1 is a fungal protein secreted by Verticillium dahliae . Our previous researches showed that this protein could induce hypersensitive responses-like necrosis and systemic acquired resistance (SAR) in cotton and tobacco. To understand immune activation mechanisms whereby PevD1 elicits defense response, the yeast two-hybrid (Y2H) assay was performed to explore interacting protein of PevD1 in Arabidopsis thaliana , and a partner AtNRP (At5g42050) was identified. Here, AtNRP homolog in Nicotiana benthamiana was identified and designated as Nbnrp1...
2018: Frontiers in Plant Science
Lai-Hua Liu, Teng-Fei Fan, Dong-Xue Shi, Chang-Jun Li, Ming-Jie He, Yi-Yin Chen, Lei Zhang, Chao Yang, Xiao-Yuan Cheng, Xu Chen, Di-Qin Li, Yi-Chen Sun
Although many members encoding different ammonium- and nitrate-transporters (AMTs, NRTs) were identified and functionally characterized from several plant species, little is known about molecular components for [Formula: see text]- and [Formula: see text] acquisition/transport in tobacco, which is often used as a plant model for biological studies besides its agricultural and industrial interest. We reported here the first molecular identification in tobacco ( Nicotiana tabacum ) of nine AMTs and four NRTs , which are respectively divided into four ( AMT1/2/3/4 ) and two ( NRT1/2 ) clusters and whose functionalities were preliminarily evidenced by heterologous functional-complementation in yeast or Arabidopsis...
2018: Frontiers in Plant Science
Anne A Madden, Mary Jane Epps, Tadashi Fukami, Rebecca E Irwin, John Sheppard, D Magdalena Sorger, Robert R Dunn
Many species of yeast are integral to human society. They produce many of our foods, beverages and industrial chemicals, challenge us as pathogens, and provide models for the study of our own biology. However, few species are regularly studied and much of their ecology remains unclear, hindering the development of knowledge that is needed to improve the relationships between humans and yeasts. There is increasing evidence that insects are an essential component of ascomycetous yeast ecology. We propose a 'dispersal-encounter hypothesis' whereby yeasts are dispersed by insects between ephemeral, spatially disparate sugar resources, and insects, in turn, obtain the benefits of an honest signal from yeasts for the sugar resources...
March 28, 2018: Proceedings. Biological Sciences
Dan Chen, Chao Yang, Sha Liu, Weijian Hang, Xianghong Wang, Juan Chen, Anbing Shi
Arf6/ARF-6 is a crucial regulator of the endosomal phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) pool in endocytic recycling. To further characterize ARF-6 regulation, we performed an ARF-6 interactor screen in Caenorhabditis elegans and identified SAC-1, the homologue of the phosphoinositide phosphatase Sac1p in yeast, as a novel ARF-6 partner. In the absence of ARF-6, basolateral endosomes show a loss of SAC-1 staining in epithelial cells. Steady-state cargo distribution assays revealed that loss of SAC-1 specifically affected apical secretory delivery and basolateral recycling...
March 21, 2018: Journal of Cell Biology
Matthew J Rossi, William K M Lai, B Franklin Pugh
General regulatory factors (GRFs), such as Reb1, Abf1, Rap1, Mcm1, and Cbf1, positionally organize yeast chromatin through interactions with a core consensus DNA sequence. It is assumed that sequence recognition via direct base readout suffices for specificity and that spurious nonfunctional sites are rendered inaccessible by chromatin. We tested these assumptions through genome-wide mapping of GRFs in vivo and in purified biochemical systems at near-base pair (bp) resolution using several ChIP-exo-based assays...
March 21, 2018: Genome Research
Darjan Andrejc, Matic Legiša
PURPOSE: Cancer cells consume more glucose than normal human cells and convert most glucose into lactate. It has been proposed that deregulated glycolysis is triggered by the posttranslational modification of 85 kDa muscle-type 6-phosphofructo-1-kinase (PFK-M) which is cleaved by a specific protease to form shorter, highly active, feedback-inhibition-resistant PFK-M fragments. PRINCIPAL RESULTS: To find the protease involved in PFK-M modification, analyses of the protease target sites on the human PFK-M enzyme yielding 45-47 kDa fragments were performed in silico...
March 18, 2018: Biochimica et Biophysica Acta
Alyssa Flobinus, Nicolas Chevigny, Phillida A Charley, Tanja Seissler, Elodie Klein, Claudine Bleykasten-Grosshans, Claudio Ratti, Salah Bouzoubaa, Jeffrey Wilusz, David Gilmer
The RNA3 species of the beet necrotic yellow vein virus (BNYVV), a multipartite positive-stranded RNA phytovirus, contains the 'core' nucleotide sequence required for its systemic movement in Beta macrocarpa . Within this 'core' sequence resides a conserved "coremin" motif of 20 nucleotides that is absolutely essential for long-distance movement. RNA3 undergoes processing steps to yield a noncoding RNA3 (ncRNA3) possessing "coremin" at its 5' end, a mandatory element for ncRNA3 accumulation...
March 19, 2018: Viruses
Matthew W McCarthy, Thomas J Walsh
Discovering new drugs for treatment of invasive fungal infections is an enduring challenge. There are only three major classes of antifungal agents, and no new class has been introduced into clinical practice in more than a decade. However, recent advances in our understanding of the fungal life cycle, functional genomics, proteomics, and gene mapping have enabled the identification of new drug targets to treat these potentially deadly infections. In this paper, we examine amino acid transport mechanisms and metabolism as potential drug targets to treat invasive fungal infections, including pathogenic yeasts, such as species of Candida and Cryptococcus , as well as molds, such as Aspergillus fumigatus ...
March 19, 2018: International Journal of Molecular Sciences
Rachel L Vannette, Tadashi Fukami
Background and Aims: Flowers can be highly variable in nectar volume and chemical composition, even within the same plant, but the causes of this variation are not fully understood. One potential cause is nectar-colonizing bacteria and yeasts, but experimental tests isolating their effects on wildflowers are largely lacking. This study examines the effects of dominant species of yeasts and bacteria on the hummingbird-pollinated shrub, Mimulus aurantiacus, in California. Methods: Wildflowers were inoculated with field-relevant titres of either the yeast Metschnikowia reukaufii or the bacterium Neokomagataea sp...
March 17, 2018: Annals of Botany
Wenhui Jiang, Tianxiang Liu, Wenzhi Nan, Diddugodage Chamila Jeewani, Yanlu Niu, Chunlian Li, Yong Wang, Xue Shi, Cong Wang, Jiahuan Wang, Yang Li, Xin Gao, Zhonghua Wang
The purple pericarp of bread wheat (Triticum aestivum L.) is a useful source of dietary anthocyanins. Previous mapping results indicated that the purple pericarp is controlled by two complementary genes located on chromosomes 7D and 2A. However, the related genes and their effects on regulating the purple pericarp are unknown. In this study, two transcription factors were characterised as anthocyanin activators in purple pericarp: TaPpm1 (purple pericarp-MYB 1) and TaPpb1 (purple pericarp-bHLH 1). Three nonfunctional variants were detected in the coding sequence of TaPpm1 from non-purple seed lines, in which the function of TaPpm1 was destroyed by insertion-caused frame shift or truncated peptides...
March 17, 2018: Journal of Experimental Botany
Lisa M Tuttle, Derek Pacheco, Linda Warfield, Jie Luo, Jeff Ranish, Steven Hahn, Rachel E Klevit
Transcription activation domains (ADs) are inherently disordered proteins that often target multiple coactivator complexes, but the specificity of these interactions is not understood. Efficient transcription activation by yeast Gcn4 requires its tandem ADs and four activator-binding domains (ABDs) on its target, the Mediator subunit Med15. Multiple ABDs are a common feature of coactivator complexes. We find that the large Gcn4-Med15 complex is heterogeneous and contains nearly all possible AD-ABD interactions...
March 20, 2018: Cell Reports
Xu Zheng, Ali Beyzavi, Joanna Krakowiak, Nikit Patel, Ahmad S Khalil, David Pincus
Clonal populations of cells exhibit cell-to-cell variation in the transcription of individual genes. In addition to this noise in gene expression, heterogeneity in the proteome and the proteostasis network expands the phenotypic diversity of a population. Heat shock factor 1 (Hsf1) regulates chaperone gene expression, thereby coupling transcriptional noise to proteostasis. Here we show that cell-to-cell variation in Hsf1 activity is an important determinant of phenotypic plasticity. Budding yeast cells with high Hsf1 activity were enriched for the ability to acquire resistance to an antifungal drug, and this enrichment depended on Hsp90, a known phenotypic capacitor and canonical Hsf1 target...
March 20, 2018: Cell Reports
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