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Yeast two hybrid

Gizem Caliskan, Ikbal C Baris, Ferhan Ayaydin, Melanie J Dobson, Muge Senarisoy, Imre M Boros, Zeki Topcu, Sevil Zencir
General Control Non-derepressible 5 (GCN5) and Alteration/Deficiency in Activation 2 and 3 proteins (ADA2 and ADA3, respectively) are subunits of the Histone AcetylTransferase (HAT) module of SAGA- and ATAC-type co-activators. We previously reported four new interacting partners of human ADA3 identified by screening a human fetal brain cDNA library using yeast two hybrid technology. One of these partners was Apoptosis-Antagonizing Transcription Factor (AATF), also known as Che-1, an RNA polymerase II-binding protein with a number of roles in different cellular processes including regulation of transcription, cell proliferation, cell cycle control, DNA damage responses and apoptosis...
2017: PloS One
Gaofeng Zhang, Tingting Lu, Wenwen Miao, Lirong Sun, Mi Tian, Ji Wang, Fushun Hao
Abscisic acid (ABA) receptor pyrabactin resistance1/PYR1-like/regulatory components of ABA receptor (PYR1/PYL/RCAR) (named PYLs for simplicity) are core regulators of ABA signaling, and have been well studied in Arabidopsis and rice. However, knowledge is limited about the PYL family regarding genome organization, gene structure, phylogenesis, gene expression and protein interaction with downstream targets in Gossypium. A comprehensive analysis of the Gossypium PYL family was carried out, and 21, 20, 40 and 39 PYL genes were identified in the genomes from the diploid progenitor G...
2017: PeerJ
Fanglin Zheng, Yanli Cao, Lei Wang, Xinxing Lv, Xiangfeng Meng, Weixin Zhang, Guanjun Chen, Weifeng Liu
Cellulase production in the model cellulolytic fungus Trichoderma reesei is subject to a variety of environmental and physiological conditions involving an intricate regulatory network with multiple transcription factors. Here, we identified the mating type locus protein MAT1-2-1 as an interacting partner for the key transcriptional activator Xyr1 of T. reesei cellulase genes. Yeast two-hybrid and GST pulldown analyses revealed that MAT1-2-1 directly interacted with the putative transcription activation domain (AD, 767~940 aa) and the middle homology region (MHR2, 314~632 aa) of Xyr1...
December 11, 2017: Scientific Reports
Xiaoyan Cui, Lu Lu, Ying Wang, Xingxing Yuan, Xin Chen
Soybean mosaic virus (SMV), a member of the Potyvirus genus, is a prevalent and devastating viral pathogen in soybean-growing regions worldwide. Potyvirus replication occurs in the 6K2-induced viral replication complex at endoplasmic reticulum exit sites. Potyvirus-encoded P3 is also associated with the endoplasmic reticulum and is as an essential component of the viral replication complex, playing a key role in viral replication. This study provides evidence that the soybean (Glycine max) reticulon homology domain protein (designated as GmRHP) interacts with SMV P3 by using a two-hybrid yeast system to screen a soybean cDNA library...
December 8, 2017: Biochemical and Biophysical Research Communications
Yan Wang, Tingting Li, Shanita Judith John, Mingjie Chen, Junli Chang, Guangxiao Yang, Guangyuan He
Drought is one of the major environmental stresses to plants. The calcium sensor, calcineurin B-like (CBL) proteins, and their interacting protein kinases (CIPK) play important roles in responding to abiotic stresses. In this study, we functionally characterized a CIPK gene from Triticum aestivum designated TaCIPK27. The transcriptional levels of TaCIPK27 were increased both in roots and leaves after treatment with polyethylene glycol 8000, abscisic acid and H2O2. Besides, TaCIPK27 interacted with AtCBL1, AtCBL3, AtCBL4, AtCBL5 and AtCBL9 in yeast two-hybrid assays...
December 1, 2017: Plant Physiology and Biochemistry: PPB
Rui Zhou, Ziyun Shao, Jian Liu, Wanqi Zhan, Qingzu Gao, Zhihua Pan, Ling Wu, Lijun Xu, Yanqing Ding, Liang Zhao
Overexpression of LIM and SH3 protein 1 (LASP1) is required for colorectal cancer (CRC) development and progression. Here, C-Jun activation domain-binding protein-1 (Jab1), also known as COPS5, was verified as a new LASP1-interacting protein through yeast two-hybrid assay. The role of COPS5 in LASP1-mediated CRC progression remains unknown. GST pull-down assay indicated that the SH3 domain of LASP1 could directly bind to MPN domain of COPS5. In vitro gain- and loss-of-function analyses revealed the stimulatory role of COPS5 on CRC cell proliferation, migration and invasion...
December 11, 2017: International Journal of Cancer. Journal International du Cancer
Huisheng Liu, Qiao Xue, Qiaoying Zeng, Zixiang Zhu, Haixue Zheng
Foot-and-mouth disease virus (FMDV) is the etiological agent of FMD, which affects domestic and wild cloven-hoofed animals. The structural protein VP1 plays an important role in FMDV pathogenesis. However, the interacting partners of VP1 in host cells and the effects of these interactions in FMDV replication remain incompletely elucidated. Here, we identified a porcine cell protein, serine/threonine kinase 3 (STK3), which interacts with FMDV VP1 using the yeast two-hybrid system. The VP1-STK3 interaction was further confirmed by coimmunoprecipitation experiments in human embryonic kidney 293T and porcine kidney 15 (PK-15) cells...
2017: BioMed Research International
Zhiyang Liu, Xuejuan Li, Feng Sun, Tong Zhou, Yijun Zhou
Rice stripe virus (RSV) causes a severe disease in Oryza sativa (rice) in many Eastern Asian countries. The NS3 protein of RSV is a viral suppressor of RNA silencing, but plant host factors interacting with NS3 have not been reported yet. Here, we present evidence that expression of RSV NS3 in Arabidopsis thaliana causes developmental abnormalities. Through yeast two-hybrid screening and a luciferase complementation imaging assay, we demonstrate that RSV NS3 interacted with OsCIPK30, a CBL (calcineurin B-like proteins)-interaction protein kinase protein...
2017: Frontiers in Microbiology
Tomohiko Makiyama, Satoru Higashi, Hiroshi Sakane, Satoru Nogami, Hiromichi Shirataki
Never in mitosis A-related kinase 2A (Nek2A), a centrosomal serine/threonine kinase, is involved in mitotic progression by regulating the centrosome cycle. Particularly, Nek2A is necessary for dissolution of the intercentriole linkage between the duplicated centrosomes prior to mitosis. Nek2A activity roughly parallels its cell cycle-dependent expression levels, but the precise mechanism regulating its activity remains unclear. In this study, we found that γ-taxilin co-localized with Nek2A at the centrosome during interphase and interacted with Nek2A in yeast two-hybrid and pull-down assays and that γ-taxilin regulated centrosome disjunction in a Nek2A-dependent manner...
December 7, 2017: Experimental Cell Research
Chi Zhang, Huw A Ogilvie, Alexei J Drummond, Tanja Stadler
Reticulate species evolution, such as hybridization or introgression, is relatively common in nature. In the presence of reticulation, species relationships can be captured by a rooted phylogenetic network, and orthologous gene evolution can be modeled as bifurcating gene trees embedded in the species network. We present a Bayesian approach to jointly infer species networks and gene trees from multilocus sequence data. A novel birthhybridization process is used as the prior for the species network, and we assume a multispecies network coalescent (MSNC) prior for the embedded gene trees...
December 6, 2017: Molecular Biology and Evolution
Yasmine Ligerot, Alexandre de Saint Germain, Tanya Waldie, Christelle Troadec, Sylvie Citerne, Nikita Kadakia, Jean-Paul Pillot, Michael Prigge, Grégoire Aubert, Abdelhafid Bendahmane, Ottoline Leyser, Mark Estelle, Frédéric Debellé, Catherine Rameau
Strigolactones (SLs) are well known for their role in repressing shoot branching. In pea, increased transcript levels of SL biosynthesis genes are observed in stems of highly branched SL deficient (ramosus1 (rms1) and rms5) and SL response (rms3 and rms4) mutants indicative of negative feedback control. In contrast, the highly branched rms2 mutant has reduced transcript levels of SL biosynthesis genes. Grafting studies and hormone quantification led to a model where RMS2 mediates a shoot-to-root feedback signal that regulates both SL biosynthesis gene transcript levels and xylem sap levels of cytokinin exported from roots...
December 8, 2017: PLoS Genetics
Qiaofeng Yang, Sihong Sang, Yao Chen, Zhaoyun Wei, Peng Wang
Seed germination and subsequent seedling development are critical phases in plant. These processes are regulated by a complex molecular network in which sugar has been reported to play an essential role. However, factors affecting sugar responses remain to be fully elucidated. In this study, we demonstrate that AtIPK2β, known to participate in the synthesis of myo-inositol 1,2,3,4,5,6-hexakisphosphate (IP6, phytate), affects Arabidopsis responses to glucose during seed germination. Loss-of-function mutant atipk2β showed increased sensitivity to 6% glucose and paclobutrazol (PAC)...
December 4, 2017: Plant & Cell Physiology
Arthur R Gorter de Vries, Philip A de Groot, Marcel van den Broek, Jean-Marc G Daran
BACKGROUND: The ease of use of CRISPR-Cas9 reprogramming, its high efficacy, and its multiplexing capabilities have brought this technology at the forefront of genome editing techniques. Saccharomyces pastorianus is an aneuploid interspecific hybrid of Saccharomyces cerevisiae and Saccharomyces eubayanus that has been domesticated for centuries and is used for the industrial fermentation of lager beer. For yet uncharacterised reasons, this hybrid yeast is far more resilient to genetic alteration than its ancestor S...
December 5, 2017: Microbial Cell Factories
Kristoffer Krogerus, Sami Holmström, Brian Gibson
Interspecific hybridization is a valuable tool for developing and improving brewing yeast in a number of industry-relevant aspects. However, the genomes of newly formed hybrids can be unstable. Here, we exploited this trait by adapting four brewing yeast strains, three of which were de novo interspecific lager hybrids with different ploidy levels, to high ethanol concentrations in an attempt to generate variant strains with improved fermentation performance in high-gravity wort. Through a batch fermentation-based adaptation process and selection based on a two-step screening process, we obtained eight variant strains which we compared to the wild-type strains in 2L-scale wort fermentations replicating industrial conditions...
December 1, 2017: Applied and Environmental Microbiology
Anirudha Dixit, Parul Tomar, Evan Vaine, Hesham Abdullah, Samuel Hazen, Om Parkash Dhankher
Members of Stress Associated Protein (SAP) family in plants have been shown to impart tolerance to multiple abiotic stresses, however, their mode of action in providing tolerance to multiple abiotic stresses is largely unknown. There are 14 SAP genes in Arabidopsis thaliana containing A20, AN1, and Cys2-His2 zinc finger domains. AtSAP13, a member of the SAP family, carries two AN1 zinc finger domains and an additional Cys2-His2 domain. AtSAP13 transcripts showed upregulation in response to Cd, ABA and salt stresses...
November 30, 2017: Plant, Cell & Environment
Jingjing Huang, Adnan Khan Niazi, David Young, Leonardo Astolfi Rosado, Didier Vertommen, Nandita Bodra, Mohamed Ragab Abdelgawwad, Florence Vignols, Bo Wei, Khadija Wahni, Talaat Bashandy, Laetitia Bariat, Frank Van Breusegem, Joris Messens, Jean-Philippe Reichheld
Plant malate dehydrogenase (MDH) isoforms are found in different cell compartments and function in key metabolic pathways. It is well known that the chloroplastic NADP-dependent MDH activities are strictly redox regulated and controlled by light. However, redox dependence of other NAD-dependent MDH isoforms have been less studied. Here, we show by in vitro biochemical characterization that the major cytosolic MDH isoform (cytMDH1) is sensitive to H2O2 through sulfur oxidation of cysteines and methionines. CytMDH1 oxidation affects the kinetics, secondary structure, and thermodynamic stability of cytMDH1...
November 29, 2017: Journal of Experimental Botany
Gianluca Telesio, Ivan Scudiero, Maddalena Pizzulo, Pellegrino Mazzone, Tiziana Zotti, Serena Voccola, Immacolata Polvere, Pasquale Vito, Romania Stilo
The three CARD-containing MAGUK (CARMA) proteins function as scaffolding molecules that regulate activation of the pro-inflammatory transcription factor NF-κB. Recently, mutations in CARMA2 have been linked to psoriasis susceptibility due to their acquired altered capacity to activate NF-κB. By means of two-hybrid screening with yeast, we identified RING finger protein 7 (RNF7) as an interactor of CARMA2. We present evidence that RNF7 functions as a negative regulator of the NF-κB-activating capacity of CARMA2...
December 1, 2017: International Journal of Molecular Sciences
Ling Wang, Xue-Lian Zhang, Lu Wang, Yanan Tian, Ning Jia, Shuzhen Chen, Ning-Bo Shi, Xuemei Huang, Chu Zhou, Yaowen Yu, Zhao-Qi Zhang, Xue-Qun Pang
WRKY transcription factors (TFs) play important roles in stress responses in planta. However, the function of WRKY TFs in the regulation of fruit ripening is unclear. Here, 23 tomato SlWRKYs that are similar to ethylene-responsive WRKY genes from other plant species, or show up-regulation during fruit ripening in previous genome-wide study, were selected, and their function in fruit ripening was investigated. Twelve SlWRKYs were found to be responsive to ethylene (SlER-WRKYs), showing expression patterns similar to those of genes related to fruit ripening...
November 30, 2017: Scientific Reports
Min Woo Kim, Wenjing Wang, Mateo I Sanchez, Robert Coukos, Mark von Zastrow, Alice Y Ting
Transcriptional assays, such as yeast two-hybrid and TANGO, that convert transient protein-protein interactions (PPIs) into stable expression of transgenes are powerful tools for PPI discovery, screens, and analysis of cell populations. However, such assays often have high background and lose information about PPI dynamics. We have developed SPARK (Specific Protein Association tool giving transcriptional Readout with rapid Kinetics), in which proteolytic release of a membrane-tethered transcription factor (TF) requires both a PPI to deliver a protease proximal to its cleavage peptide and blue light to uncage the cleavage site...
November 30, 2017: ELife
Y Wang, N Li, X Zhao, J Hu, Y He, T Hu, S Wang, Y Wang, K Cao
Apple chlorotic leaf spot virus (ACLSV) movement protein (P50) is involved in cell-to-cell transport and influences the long-distance spread of silencing activity. Previously, we obtained 69 P50-interacting proteins from Malus sylvestris cv. R12740-7A and using bioinformatics analyzed their biological functions. In this study, we used the GAL4-based two-hybrid yeast system and His pull-down assays to confirm an interaction between PR-10 of M. sylvestris cv. R12740-7A and ACLSV P50. Our results provide a theoretical basis for further research on the biological function of PR-10 in ACLSV infection and the interacting mechanism between host and virus...
2017: Acta Virologica
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