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https://www.readbyqxmd.com/read/28528573/a-sensitive-semi-quantitative-mammalian-two-hybrid-assay
#1
Elisabeth Riegel, Thomas Heimbucher, Thomas Höfer, Thomas Czerny
Protein-protein interactions critically determine the function of a protein within the cell. Several methods have been developed for the analysis of protein interactions, including two-hybrid assays in yeast and mammals. Mammalian two-hybrid systems provide the ideal physiological environment to study the interactions of mammalian proteins; however, these approaches are limited in sensitivity and their ability to quantify interaction strength. Here, we present an inducible mammalian two-hybrid (iM2H) system using the small-molecule dimerizer rapalog for recruitment of multiple transactivation domains into the M2H system...
May 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/28527764/differential-stress-response-of-saccharomyces-hybrids-revealed-by-monitoring-hsp104-aggregation-and-disaggregation
#2
Claudia Kempf, Klaus Lengeler, Jürgen Wendland
Proteotoxic stress may occur upon exposure of yeast cells to different stress conditions. The induction of stress response mechanisms is important for cells to adapt to changes in the environment and ensure survival. For example, during exposure to elevated temperatures the expression of heat shock proteins such as Hsp104 is induced in yeast. Hsp104 extracts misfolded proteins from aggregates to promote their refolding. We used an Hsp104-GFP reporter to analyze the stress profiles of Saccharomyces species hybrids...
July 2017: Microbiological Research
https://www.readbyqxmd.com/read/28527583/molecular-characterization-of-aspergillus-fumigatus-tcsc-a-characteristic-type-iii-hybrid-histidine-kinase-of-filamentous-fungi-harboring-six-hamp-domains
#3
Anja Spadinger, Frank Ebel
The type III hybrid histidine kinase (HHK) TcsC enables the pathogenic mold Aspergillus fumigatus to thrive under hyperosmotic conditions. It is, moreover, of particular interest, since it is the target of certain antifungal agents, such as fludioxonil. This study was aimed at a functional characterization of the domains that constitute the sensing and the kinase module of TcsC. The sensing module consists of six HAMP domains, an architecture that is commonly found in type III HHKs of filamentous fungi. To dissect the functional role of the individual domains, we have analyzed a set of truncated derivatives of TcsC with respect to their impact on fungal growth and their ability to respond to hyperosmotic stress and fludioxonil...
May 11, 2017: International Journal of Medical Microbiology: IJMM
https://www.readbyqxmd.com/read/28527011/the-prdm9-krab-domain-is-required-for-meiosis-and-involved-in-protein-interactions
#4
Yukiko Imai, Frédéric Baudat, Miguel Taillepierre, Marcello Stanzione, Attila Toth, Bernard de Massy
PR domain-containing protein 9 (PRDM9) is a major regulator of the localization of meiotic recombination hotspots in the human and mouse genomes. This role involves its DNA-binding domain, which is composed of a tandem array of zinc fingers, and PRDM9-dependent trimethylation of histone H3 at lysine 4. PRDM9 is a member of the PRDM family of transcription regulators, but unlike other family members, it contains a Krüppel-associated box (KRAB)-related domain that is predicted to be a potential protein interaction domain...
May 19, 2017: Chromosoma
https://www.readbyqxmd.com/read/28526529/adding-biological-meaning-to-human-protein-protein-interactions-identified-by-yeast-two-hybrid-screenings-a-guide-through-bioinformatics-tools
#5
Juliana Felgueiras, Joana Vieira Silva, Margarida Fardilha
"A man is known by the company he keeps" is a popular expression that perfectly fits proteins. A common approach to characterize the function of a target protein is to identify its interacting partners and thus infer its roles based on the known functions of the interactors. Protein-protein interaction networks (PPINs) have been created for several organisms, including humans, primarily as results of high-throughput screenings, such as yeast two-hybrid (Y2H). Their unequivocal use to understand events underlying human pathophysiology is promising in identifying genes and proteins associated with diseases...
May 16, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28525953/human-yeast-hybrids-new-visions-to-genetic-disorders-and-drug-discovery
#6
Najmeh Zarei, Vahid Khalaj
Yeast has been a very helpful organism for centuries, especially with respect to fermentation of sugars and production of bread. However, for an even longer time, yeast has been a distant relative of humans having diverged from a common ancestor, about one billion years ago. More than one third of the yeast genes have human counterparts, despite this evolutionary distance. Yeast and human orthologs perform the same or similar functions. Investigations have demonstrated that 9-92% of the amino acid sequences in similar human and yeast proteins overlap...
July 2017: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/28522455/psa3-a-protein-on-the-stromal-face-of-the-thylakoid-membrane-promotes-photosystem-i-accumulation-in-cooperation-with-the-assembly-factor-pyg7
#7
Jie Shen, Rosalind Williams-Carrier, Alice Barkan
Photosystem I (PSI) is a large protein-pigment complex located in the thylakoid membrane in cyanobacteria, plants, and algae. Although the structure and components of PSI are well characterized, mechanisms that orchestrate its assembly are poorly understood. In this study, we discovered a novel nucleus-encoded protein, Photosystem I Assembly 3 (PSA3), that is required for PSI accumulation. PSA3 is conserved among green photosynthetic eukaryotes but is lacking in cyanobacteria. Mutations in the psa3 gene cause the specific loss of PSI in Arabidopsis and maize...
May 18, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28521960/yeast-dynamin-associates-with-the-garp-tethering-complex-for-endosome-to-golgi-traffic
#8
Uma Saimani, Jared Smothers, Hyoeun McDermott, Pelin Makaraci, Kyoungtae Kim
Yeast dynamin, Vacuolar Protein Sorting 1 (Vps1), has been implicated in recycling traffic from the endosome to the trans-Golgi network (TGN). Previous research showed a genetic interaction of Vps1 with all components of the GARP tethering complex, which anchors vesicles at the late Golgi membrane. We used the yeast two-hybrid system and have identified a 33 amino acid segment of Vps51, a GARP subunit, that interacts with Vps1. Based on sequence homology between Vps51 and its mammalian homolog Ang2 in the 33 amino acids stretch, we identified two key residues of Vps51, E127 and Y129, that bind Vps1...
May 8, 2017: European Journal of Cell Biology
https://www.readbyqxmd.com/read/28508886/protein-protein-interactions-and-metabolite-channelling-in-the-plant-tricarboxylic-acid-cycle
#9
Youjun Zhang, Katherine F M Beard, Corné Swart, Susan Bergmann, Ina Krahnert, Zoran Nikoloski, Alexander Graf, R George Ratcliffe, Lee J Sweetlove, Alisdair R Fernie, Toshihiro Obata
Protein complexes of sequential metabolic enzymes, often termed metabolons, may permit direct channelling of metabolites between the enzymes, providing increased control over metabolic pathway fluxes. Experimental evidence supporting their existence in vivo remains fragmentary. In the present study, we test binary interactions of the proteins constituting the plant tricarboxylic acid (TCA) cycle. We integrate (semi-)quantitative results from affinity purification-mass spectrometry, split-luciferase and yeast-two-hybrid assays to generate a single reliability score for assessing protein-protein interactions...
May 16, 2017: Nature Communications
https://www.readbyqxmd.com/read/28506636/multiple-dna-interactions-contribute-to-the-initiation-of-telomerase-elongation
#10
Ahu Karademir Andersson, Cecilia Gustafsson, Roopesh Krishnankutty, Marita Cohn
Telomerase maintains telomere length and chromosome integrity by adding short tandem repeats of single-stranded DNA to the 3' ends, via reverse transcription of a defined template region of its RNA subunit. To further understand the telomerase elongation mechanism, we studied the primer utilization and extension activity of the telomerase from the budding yeast Naumovozyma castellii (Saccharomyces castellii), which displays a processive nucleotide and repeat addition polymerization. For the efficient initiation of canonical elongation, telomerase required 4nt primer 3' end complementarity to the template RNA...
May 12, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28505371/the-arm-domain-of-armadillo-repeat-kinesin-1-is-not-required-for-microtubule-catastrophe-but-can-negatively-regulate-nima-related-kinase-6-in-arabidopsis-thaliana
#11
Ryan C Eng, Laryssa S Halat, Samuel J Livingston, Tatsuya Sakai, Hiroyasu Motose, Geoffrey Wasteneys
Microtubules are dynamic filaments, the assembly and disassembly of which are under precise control of various associated proteins, including motor proteins and regulatory enzymes. In Arabidopsis thaliana, two such proteins are the ARMADILLO-REPEAT KINESIN 1 (ARK1), which promotes microtubule disassembly, and the NIMA-RELATED KINASE 6 (NEK6), which has a role in organizing microtubule arrays. Previous yeast 2-hybrid and in vitro pull-down assays determined that NEK6 can interact with ARK1 through the latter protein's Armadillo-repeat (ARM) cargo domain...
May 13, 2017: Plant & Cell Physiology
https://www.readbyqxmd.com/read/28494021/rice-black-streaked-dwarf-virus-p7-2-forms-a-scf-complex-through-binding-to-oryza-sativa-skp1-like-proteins-and-interacts-with-gid2-involved-in-the-gibberellin-pathway
#12
Tao Tao, Cui-Ji Zhou, Qian Wang, Xiang-Ru Chen, Qian Sun, Tian-Yu Zhao, Jian-Chun Ye, Ying Wang, Zong-Ying Zhang, Yong-Liang Zhang, Ze-Jian Guo, Xian-Bing Wang, Da-Wei Li, Jia-Lin Yu, Cheng-Gui Han
As a core subunit of the SCF complex that promotes protein degradation through the 26S proteasome, S-phase kinase-associated protein 1 (SKP1) plays important roles in multiple cellular processes in eukaryotes, including gibberellin (GA), jasmonate, ethylene, auxin and light responses. P7-2 encoded by Rice black streaked dwarf virus (RBSDV), a devastating viral pathogen that causes severe symptoms in infected plants, interacts with SKP1 from different plants. However, whether RBSDV P7-2 forms a SCF complex and targets host proteins is poorly understood...
2017: PloS One
https://www.readbyqxmd.com/read/28486617/vppub24-a-novel-gene-from-chinese-grapevine-vitis-pseudoreticulata-targets-vpice1-to-enhance-cold-tolerance
#13
Wenkong Yao, Lei Wang, Jie Wang, Fuli Ma, Yazhou Yang, Chen Wang, Weihuo Tong, Jianxia Zhang, Yan Xu, Xiping Wang, Chaohong Zhang, Yuejin Wang
The ubiquitination system plays important roles in the degradation and modification of substrate proteins. In this study, we characterize a putative U-box type E3 ubiquitin ligase gene, VpPUB24 (plant U-box protein 24), from Chinese wild grapevine, Vitis pseudoreticulata accession Baihe-35-1. We show that VpPUB24 is induced by a number of stresses, especially cold treatment. Real-time PCR analysis indicated that the PUB24 transcripts were increased after cold stress in different grapevine species, although the relative expression level was different...
May 9, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28483877/lnk1-and-lnk2-co-repressors-interact-with-the-myb3-transcription-factor-in-phenylpropanoid-biosynthesis
#14
Meiliang Zhou, Kaixuan Zhang, Zhanmin Sun, Mingli Yan, Cheng Chen, Xinquan Zhang, Yixiong Tang, Yanmin Wu
Subgroup 4 of R2R3-MYB transcription factors (TFs) consist of four members, MYB3, MYB4, MYB7 and MYB32, which possess the conserved EAR repression motif (pdLHLD/LLxiG/S) in their C-termini. Here, we show that MYB3 is a newly identified repressor in Arabidopsis phenylpropanoids biosynthesis. However, the repression mechanism of MYB3 is completely different from MYB4, MYB7 and MYB32. Yeast two hybrid screening using MYB3 as a bait isolates NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED1 (LNK1) and LNK2, members of a small family of four LNK proteins...
May 8, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28483053/gm1-mmp-is-involved-in-growth-and-development-of-leaf-and-seed-and-enhances-tolerance-to-high-temperature-and-humidity-stress-in-transgenic-arabidopsis
#15
Sushuang Liu, Yanmin Liu, Yanhong Jia, Jiaping Wei, Shuang Wang, Xiaolin Liu, Yali Zhou, Yajing Zhu, Weihong Gu, Hao Ma
Matrix metalloproteinases (MMPs) are a family of zinc- and calcium-dependent endopeptidases. Gm1-MMP was found to play an important role in soybean tissue remodeling during leaf expansion. In this study, Gm1-MMP was isolated and characterized. Its encoding protein had a relatively low phylogenetic relationship with the MMPs in other plant species. Subcellular localization indicated that Gm1-MMP was a plasma membrane protein. Gm1-MMP showed higher expression levels in mature leaves, old leaves, pods, and mature seeds, as well as was involved in the development of soybean seed...
June 2017: Plant Science: An International Journal of Experimental Plant Biology
https://www.readbyqxmd.com/read/28476868/a-new-method-reverse-yeast-two-hybrid-array-rytha-identifies-mutants-that-dissociate-the-physical-interaction-between-elg1-and-slx5
#16
Ifat Lev, Keren Shemesh, Marina Volpe, Soumitra Sau, Nelly Levinton, Maya Molco, Shivani Singh, Batia Liefshitz, Shay Ben Aroya, Martin Kupiec
The vast majority of processes within the cell are carried out by proteins working in conjunction. The Yeast Two Hybrid (Y2H) methodology allows the detection of physical interactions between any two interacting proteins. Here we describe a novel systematic genetic methodology: "Reverse Yeast Two Hybrid Array (RYTHA)," that allows the identification of proteins required for modulating the physical interaction between two given proteins. Our assay starts with a yeast strain in which the physical interaction of interest can be detected by growth on media lacking histidine, in the context of the Yeast Two Hybrid (Y2H) methodology...
May 5, 2017: Genetics
https://www.readbyqxmd.com/read/28475733/quatre-quart1-is-an-indispensable-u12-intron-containing-gene-that-plays-a-crucial-role-in-arabidopsis-development
#17
Kyung Jin Kwak, Bo Mi Kim, Kwanuk Lee, Hunseung Kang
Despite increasing understanding of the importance of the splicing of U12-type introns in plant development, the key question of which U12 intron-containing genes are essential for plant development has not yet been explored. Here, we assessed the functional role of the quatre-quart1 (QQT1) gene, one of the ~230 U12 intron-containing genes in Arabidopsis thaliana. Expression of QQT1 in the U11/U12-31K small nuclear ribonucleoprotein mutant (31k) rescued the developmental-defect phenotypes of the 31k mutant, whereas the miRNA-mediated qqt1 knockdown mutants displayed severe defects in growth and development, including severely arrested stem growth, small size, and the formation of serrated leaves...
May 5, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28472636/near-neighbor-interactions-of-the-membrane-embedded-subunits-of-the-mitochondrial-atp-synthase-of-a-chlorophycean-alga
#18
Lorenzo Sánchez-Vásquez, Miriam Vázquez-Acevedo, Javier de la Mora, Félix Vega-deLuna, Pierre Cardol, Claire Remacle, Georges Dreyfus, Diego González-Halphen
Mitochondrial F1FO-ATP synthase of the chlorophycean algae Polytomella sp. can be isolated as a highly stable dimeric complex of 1600kDa. It is composed of eight highly conserved orthodox subunits (α, β, γ, δ, ε, OSCP, a, and c) and nine subunits (Asa1-9) that are exclusive of chlorophycean algae. The Asa subunits replace those that build up the peripheral stalk and the dimerization domains of the ATP synthase in other organisms. Little is known about the disposition of subunits Asa6, Asa8 and Asa9, that are predicted to have transmembrane stretches and that along with subunit a and a ring of c-subunits, seem to constitute the membrane-embedded Fo domain of the algal ATP synthase...
May 1, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28472300/interaction-of-the-mineralocorticoid-receptor-with-rack1-and-its-role-in-aldosterone-signaling
#19
Maniselvan Kuppusamy, Elise P Gomez-Sanchez, Lauren N Beloate, Maria Plonczynski, Aniko Naray-Fejes-Toth, Geza Fejes-Toth, Celso E Gomez-Sanchez
The mineralocorticoid receptor (MR) is a member of the steroid-thyroid hormone receptor superfamily of ligand-dependent transcription factors with diverse functions including the biological actions of aldosterone. Identification of the various transcriptional co-regulators of MR is essential for understanding the complexity of MR signaling pathways under physiological and pathological conditions. We used a yeast two-hybrid system to find proteins that interact with a full-length MR and found, among other proteins, that MR interacted specifically with Receptor for Activated C Kinase 1 (RACK1), a scaffolding protein...
May 2, 2017: Endocrinology
https://www.readbyqxmd.com/read/28470426/the-yeasts-phosphorelay-systems-a-comparative-view
#20
REVIEW
Griselda Salas-Delgado, Laura Ongay-Larios, Laura Kawasaki-Watanabe, Imelda López-Villaseñor, Roberto Coria
Cells contain signal transduction pathways that mediate communication between the extracellular environment and the cell interior. These pathways control transcriptional programs and posttranscriptional processes that modify cell metabolism in order to maintain homeostasis. One type of these signal transduction systems are the so-called Two Component Systems (TCS), which conduct the transfer of phosphate groups between specific and conserved histidine and aspartate residues present in at least two proteins; the first protein is a sensor kinase which autophosphorylates a histidine residue in response to a stimulus, this phosphate is then transferred to an aspartic residue located in a response regulator protein...
June 2017: World Journal of Microbiology & Biotechnology
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