scholarly journals Interaction of porcine circovirus-like virus P1 capsid protein with host proteins

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Libin Wen ◽  
Jiaping Zhu ◽  
Fengxi Zhang ◽  
Qi Xiao ◽  
Jianping Xie ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Underlying Mechanism ◽  
Yeast Cells ◽  
Cellular Proteins ◽  
Porcine Circovirus ◽  
Yeast Two Hybrid ◽  
Host Proteins ◽  
Host Interactions ◽  
Wasting Syndrome ◽  
Two Hybrid

Abstract Background Porcine circovirus-like virus P1 is a relatively new kind of virus that is closely related to the post-weaning multisystemic wasting syndrome, congenital tremors, and abortions in swine. The molecular mechanisms of P1 virus infection and pathogenesis are fully unknown. To analyze P1 and its host interactions, we used a yeast two-hybrid (Y2H) assay to identify cellular proteins interacting with the Cap of the P1 virus. In this study, the Cap of the P1 virus exhibited no self-activation and toxicity to yeast cells and was used as bait to screen the Y2H library prepared from the pancreas tissue. Results Five cellular proteins (EEP, Ral GDS, Bcl-2-L-12, CPS1, and one not identified) were found to interact with P1 Cap. The interaction between Cap and Ral GDS was confirmed by co-immunoprecipitation. Conclusions Our data are likely to support the future investigation of the underlying mechanism of P1 infection and pathogenesis.

scholarly journals Hantavirus nucleocapsid protein interacts with the Fas-mediated apoptosis enhancer Daxx

2002 ◽  
Vol 83 (4) ◽  
pp. 759-766 ◽  
Author(s):  
Xiao-Dong Li ◽  
Tomi P. Mäkelä ◽  
Deyin Guo ◽  
Rabah Soliymani ◽  
Vesa Koistinen ◽  
...  
Keyword(s):  
Binding Sites ◽  
Nuclear Localization Signal ◽  
Nucleocapsid Protein ◽  
Molecular Mechanisms ◽  
Cellular Proteins ◽  
Yeast Two Hybrid ◽  
Localization Signal ◽  
Carboxyl Terminal ◽  
Two Hybrid ◽  
Hybrid Screening

Hantaviruses cause two severe diseases, haemorrhagic fever with renal syndrome in Eurasia and hantavirus pulmonary syndrome in the Americas. To understand more about the molecular mechanisms that lead to these diseases, the associations of Puumala virus nucleocapsid protein (PUUV-N) with cellular proteins were studied by yeast two-hybrid screening. Daxx, known as an apoptosis enhancer, was identified from a HeLa cDNA library and its interaction with PUUV-N was confirmed by GST pull-down assay, co-immunoprecipitation and co-localization studies. Furthermore, domains of interaction were mapped to the carboxyl-terminal region of 142 amino acids in Daxx and the carboxyl-terminal 57 residues in PUUV-N, respectively. In pepscan assays, the binding sites of Daxx to PUUV-N were mapped further to two lysine-rich regions, of which one overlaps the sequence of the predicted nuclear localization signal of Daxx. These data suggest a direct link between host cell machinery and a hantavirus structural component.

scholarly journals Yeast two-hybrid screening for proteins that interact with PFT in wheat

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yi He ◽  
Lei Wu ◽  
Xiang Liu ◽  
Xu Zhang ◽  
Peng Jiang ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Molecular Mechanisms ◽  
Reduction Process ◽  
Resistance Breeding ◽  
Yeast Cells ◽  
Yeast Two Hybrid ◽  
Head Blight ◽  
Fhb Resistance ◽  
Trait Locus ◽  
Two Hybrid

Abstract Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Fhb1 is the most consistently reported quantitative trait locus (QTL) for FHB resistance breeding. A pore-forming toxin-like (PFT) gene at Fhb1 was first cloned by map-based cloning and found to confer FHB resistance in wheat. Proteins often interact with each other to execute their functions. Characterization of the proteins interacting with PFT might therefore provide information on the molecular mechanisms of PFT functions. In this study, a high-quality yeast two-hybrid (Y2H) library using RNA extracted from Fusarium graminearum (Fg)-infected wheat spikes of Sumai 3 was constructed. The agglutinin domains of PFT exhibited no self-activation and toxicity to yeast cells and were used as bait to screen the Y2H library. Twenty-three proteins that interact with PFT were obtained, which were mainly involved in the ubiquitination process, clathrin coat assembly, the oxidation-reduction process, and protein phosphorylation. The expression pattern of these interacting genes was analyzed by quantitative real-time PCR. This study clarifies the protein interactions of PFT and raises a regulatory network for PFT regarding FHB resistance in wheat.

scholarly journals Implication of the Whitefly Protein Vps Twenty Associated 1 (Vta1) in the Transmission of Cotton Leaf Curl Multan Virus

2021 ◽  
Vol 9 (2) ◽  
pp. 304
Author(s):  
Yao Chi ◽  
Li-Long Pan ◽  
Shu-Sheng Liu ◽  
Shahid Mansoor ◽  
Xiao-Wei Wang
Keyword(s):  
Coat Protein ◽  
Molecular Mechanisms ◽  
Cotton Leaf ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Vacuolar Protein ◽  
Leaf Curl Disease ◽  
Two Hybrid ◽  
Asia Ii 1 ◽  
Leaf Curl

Cotton leaf curl Multan virus (CLCuMuV) is one of the major casual agents of cotton leaf curl disease. Previous studies show that two indigenous whitefly species of the Bemisia tabaci complex, Asia II 1 and Asia II 7, are able to transmit CLCuMuV, but the molecular mechanisms underlying the transmission are poorly known. In this study, we attempted to identify the whitefly proteins involved in CLCuMuV transmission. First, using a yeast two-hybrid system, we identified 54 candidate proteins of Asia II 1 that putatively can interact with the coat protein of CLCuMuV. Second, we examined interactions between the CLCuMuV coat protein and several whitefly proteins, including vacuolar protein sorting-associated protein (Vps) twenty associated 1 (Vta1). Third, using RNA interference, we found that Vta1 positively regulated CLCuMuV acquisition and transmission by the Asia II 1 whitefly. In addition, we showed that the interaction between the CLCuMuV coat protein and Vta1 from the whitefly Middle East-Asia Minor (MEAM1), a poor vector of CLCuMuV, was much weaker than that between Asia II 1 Vta1 and the CLCuMuV coat protein. Silencing of Vta1 in MEAM1 did not affect the quantity of CLCuMuV acquired by the whitefly. Taken together, our results suggest that Vta1 may play an important role in the transmission of CLCuMuV by the whitefly.

scholarly journals iTRAQ analysis of Singapore grouper iridovirus infection in a grouper embryonic cell line

2008 ◽  
Vol 89 (11) ◽  
pp. 2869-2876 ◽  
Author(s):  
Li Ming Chen ◽  
Bich Ngoc Tran ◽  
Qingsong Lin ◽  
Teck Kwang Lim ◽  
Fan Wang ◽  
...  
Keyword(s):  
Viral Infection ◽  
Cell Line ◽  
Molecular Mechanisms ◽  
Embryonic Cell ◽  
Host Proteins ◽  
Host Interactions ◽  
Embryonic Cell Line ◽  
Itraq Analysis ◽  
Liquid Chromatography Tandem Mass ◽  
First Time

We report, here, the first proteomics study of a grouper embryonic cell line (GEC) infected by Singapore grouper iridovirus (SGIV). The differential proteomes of GEC with and without viral infection were studied and quantified with iTRAQ labelling followed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Forty-nine viral proteins were recognized, of which 11 were identified for the first time. Moreover, 743 host proteins were revealed and classified into 218 unique protein groups. Fourteen host proteins were upregulated and five host proteins were downregulated upon viral infection. The iTRAQ analysis of SGIV infection in GEC provides an insight to viral and host gene products at the protein level. This should facilitate further study and the understanding of virus–host interactions, molecular mechanisms of viral infection and pathogenesis.

A set of host proteins interacting with papaya ringspot virus NIa-Pro protein identified in a yeast two-hybrid system

2012 ◽  
Vol 56 (01) ◽  
pp. 25-30 ◽  
Author(s):  
L. GAO ◽  
W. T. SHEN ◽  
P. YAN ◽  
D. C. TUO ◽  
X. Y. LI ◽  
...  
Keyword(s):  
Hybrid System ◽  
Ringspot Virus ◽  
Papaya Ringspot Virus ◽  
Yeast Two Hybrid ◽  
Host Proteins ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

scholarly journals Host targets of effectors of the lettuce downy mildew Bremia lactucae from cDNA-based yeast two-hybrid screening

2019 ◽  
Author(s):  
Alexandra J.E. Pelgrom ◽  
Claudia-Nicole Meisrimler ◽  
Joyce Elberse ◽  
Thijs Koorman ◽  
Mike Boxem ◽  
...  
Keyword(s):  
Downy Mildew ◽  
Pathogenic Bacteria ◽  
Plant Pathogens ◽  
Molecular Mechanisms ◽  
Effector Proteins ◽  
Bremia Lactucae ◽  
Yeast Two Hybrid ◽  
Target Proteins ◽  
Successful Infection ◽  
Two Hybrid

AbstractPlant pathogenic bacteria, fungi and oomycetes secrete effector proteins to manipulate host cell processes to establish a successful infection. Over the last decade the genomes and transcriptomes of many agriculturally important plant pathogens have been sequenced and vast candidate effector repertoires were identified using bioinformatic analyses. Elucidating the contribution of individual effectors to pathogenicity is the next major hurdle. To advance our understanding of the molecular mechanisms underlying lettuce susceptibility to the downy mildew Bremia lactucae, we mapped a network of physical interactions between B. lactucae effectors and lettuce target proteins. Using a lettuce cDNA library-based yeast-two-hybrid system, 61 protein-protein interactions were identified, involving 21 B. lactucae effectors and 46 unique lettuce proteins. The top ten targets based on the number of independent colonies identified in the Y2H and two targets that belong to gene families involved in plant immunity, were further characterized. We determined the subcellular localization of the fluorescently tagged target proteins and their interacting effectors. Importantly, relocalization of effectors or targets to the nucleus was observed for four effector-target pairs upon their co-expression, supporting their interaction in planta.

scholarly journals Sp100 interacts with phage ΦC31 integrase to inhibit its recombination activity.

2011 ◽  
Vol 58 (1) ◽  
Author(s):  
Yun Lin ◽  
Zhi-hui Li ◽  
Jing-Jing Wang ◽  
Gua-lan Xu ◽  
Qi Shen ◽  
...  
Keyword(s):  
Hek293 Cell ◽  
Fetal Brain ◽  
Cell Extract ◽  
Cellular Proteins ◽  
Yeast Two Hybrid ◽  
Potential Vector ◽  
Yeast Two Hybrid System ◽  
Φc31 Integrase ◽  
Brain Cdna Library ◽  
Two Hybrid

Phage ΦC31 integrase is a potential vector for the insertion of therapeutic genes into specific sites in the human genome. To understand the mechanism involved in ΦC31 integrase-mediated recombination, it is important to understand the interaction between the integrase and cellular proteins. Using a yeast two-hybrid system with pLexA-ΦC31 integrase as bait, we screened a pB42AD human fetal brain cDNA library for potential interacting cellular proteins. From the 10⁶ independent clones that were screened, 11 potential interacting clones were isolated, of which one encoded C-terminal fragment of Sp100. The interaction between Sp100 and ΦC31 integrase was further confirmed by yeast mating and co-immunoprecipitation assays. The hybridization between a ΦC31 integrase peptide array and an HEK293 cell extract revealed that residues 81RILN84 in the N-terminus of ΦC31 integrase are responsible for the interaction with Sp100. Knocking down endogenous Sp100 with Sp100-specific siRNA increased ΦC31 integrase-mediated recombination but did not impact reporter gene expression. Therefore, endogenous Sp100 may interact with ΦC31 integrase and inhibit the efficiency of ΦC31 integrase-mediated recombination.

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