scholarly journals Screening and Identification of Host Proteins Interacting with Iris lactea var. chinensis Metallothionein IlMT2a by Yeast Two-Hybrid Assay

Genes ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 554
Author(s):  
Zhiquan Wang ◽  
Longjie Ni ◽  
Liangqin Liu ◽  
Haiyan Yuan ◽  
Suzhen Huang ◽  
...  
Keyword(s):  
Stress Responses ◽  
Transgenic Arabidopsis ◽  
Biological Significance ◽  
Significant Protection ◽  
Yeast Two Hybrid ◽  
Host Proteins ◽  
Screening And Identification ◽  
First Time ◽  
Gene Regulations ◽  
Two Hybrid

Iris lactea var. chinensis (Fisch.) (I. lactea var. chinensis) is a well-known cadmium (Cd)-tolerant plant and we have previously shown that the metallothionein gene, IlMT2a, of the plant may be playing a key role in conferring the Cd tolerance. In this study, we have identified several proteins interacting with the IlMT2a by screening yeast two-hybrid library constructed from cDNAs isolated from Cd-treated I. lacteal var. chinensis plants. Putative functions of these proteins include those involved in photosynthesis, ROS scavenge, nutrient transport, and transcriptional regulation, to name a few. In particular, another metallothionein, which we assigned the name of IlMT3, was identified as an interacting partner of the IlMT2a. Unlike IlMT2a, it did not provide any significant protection against Cd toxicity in transgenic Arabidopsis thaliana L. (A. thaliana). To our knowledge, this is the first time ever reporting the interaction of two metallothionein proteins in plants. Learning the biological significance of the interaction between IlMT2a and IlMT3 would be the focus of future study and would be able to provide valuable insights into the understanding plant metallothionein’s diverse and complex roles in coordinating many important cellular physiologies including stress responses, gene regulations, and energy metabolisms.

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

Screening and Verification of Host Proteins Interacting with Neospora Caninum AMA1 Using a Yeast Two-hybrid System

2020 ◽  
Author(s):  
Baoyi Wu ◽  
Chunmei Jin ◽  
Yinan Wang ◽  
Yan Zhang ◽  
Tianyu Zheng ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Transmembrane Protein ◽  
Type I ◽  
Filamin A ◽  
Biological Functions ◽  
Yeast Two Hybrid ◽  
Apical Membrane Antigen 1 ◽  
Invasion Mechanism ◽  
First Time ◽  
Two Hybrid

Abstract Background: Neospora caninum apical membrane antigen 1 (NcAMA1) is a conservative type I transmembrane protein that is secreted by the microneme to the surface of the parasite, and is a key component of the invasion mechanism. In order to explore further the biological functions of NcAMA1 in the process of parasite invasion, we conducted research on NcAMA1 and its interacting partners. Methods: In this study, Vero cell yeast two-hybrid (Y2H) cDNA library was constructed. Using the constructed recombinant vector pGBKT7-NcAMA1 as bait, the Y2H system was used to screen the proteins that interact with NcAMA1. In addition, the interaction between NcAMA1 and the screened transmembrane emp24 domain trafficking protein 2 （Tmed2）was further verified by one-to-one Y2H experiments and pull-down, and the role of Tmed2 protein in the process of N. caninum invasion was initially verified by RNA silencing and antibody blocking experiments.Results: Our results show that, through the Y2H experiment, we have identified two proteins that interact with NcAMA1, which are the Chlorocebus sabaeus filamin A, alpha and Chlorocebus sabaeus Tmed2. When the expression of Tmed2 protein decreased or blocked, the invasion rate of N. caninum was increased.Conclusions: These findings give us a deeper understanding of the biological functions of NcAMA1, and for the first time suggest that Tmed2 may be involved in the process invasion by of N. caninum, inhibiting the invasion of parasites by interacting with the protein secreted by N. caninum.

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 A Comprehensive Arabidopsis Yeast Two-Hybrid Library for Protein-Protein Interaction Studies: A Resource to the Plant Research Community

2018 ◽  
Vol 31 (9) ◽  
pp. 899-902 ◽  
Author(s):  
Cleverson Carlos Matiolli ◽  
Maeli Melotto
Keyword(s):  
Protein Interactions ◽  
Stress Responses ◽  
Protein Function ◽  
Cdna Libraries ◽  
Optimum Conditions ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Biotic And Abiotic Stresses ◽  
Protein Protein Interaction ◽  
Two Hybrid

Yeast-two-hybrid (Y2H) cDNA library screening is a valuable tool to uncover protein-protein interactions and represents a widely used method to investigate protein function. However, low transcript representation in cDNA libraries limits the depth of the screening. We have developed a Y2H library with cDNA made from Arabidopsis leaves exposed to several stressors as well as untreated leaves. The library was built using pooled mRNA extracted from plants challenged with plant and human bacterial pathogens, the flg22 elicitor, the phytotoxin coronatine, and several hormones associated with environmental stress responses. The purpose of such a library is to maximize the discovery of protein-protein interactions that occur under optimum conditions as well as during biotic and abiotic stresses.

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