scholarly journals Identification and Characterization of an OSH1 Thiol Reductase from Populus trichocarpa

Cells ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 76
Author(s):  
Wei ◽  
Zhou ◽  
Xu ◽  
Movahedi ◽  
Sun ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Populus Trichocarpa ◽  
Expression System ◽  
Homologous Gene ◽  
Cd Stress ◽  
Ros Scavenging ◽  
Prokaryotic Expression System ◽  
Thiol Reductase ◽  
Identification And Characterization

Interferon gamma-induced lysosomal thiol reductase (GILT) is abundantly expressed in antigen-presenting cells and participates in the treatment and presentation of antigens by major histocompatibility complex II. Also, GILT catalyzes the reduction of disulfide bonds, which plays an important role in cellular immunity. (1) Background: At present, the studies of GILT have mainly focused on animals. In plants, GILT homologous gene (Arabidopsis thaliana OSH1: AtOSH1) was discovered in the forward screen of mutants with compromised responses to sulphur nutrition. However, the complete properties and functions of poplar OSH1 are unclear. In addition, CdCl2 stress is swiftly engulfing the limited land resources on which humans depend, restricting agricultural production. (2) Methods: A prokaryotic expression system was used to produce recombinant PtOSH1 protein, and Western blotting was performed to identify its activity. In addition, a simplified version of the floral-dip method was used to transform A. thaliana. (3) Results: Here, we describe the identification and characterization of OSH1 from Populus trichocarpa. The deduced PtOSH1 sequence contained CQHGX2ECX2NX4C and CXXC motifs. The transcript level of PtOSH1 was increased by cadmium (Cd) treatment. In addition, recombinant PtOSH1 reduced disulfide bonds. A stress assay showed that PtOSH1-overexpressing (OE) A. thaliana lines had greater resistance to Cd than wild-type (WT) plants. Also, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in PtOSH1-OE plants were significantly higher than those in WT A. thaliana. These results indicate that PtOSH1 likely plays an important role in the response to Cd by regulating the reactive oxygen species (ROS)-scavenging system. (4) Conclusions: PtOSH1 catalyzes the reduction of disulfide bonds and behaves as a sulfhydryl reductase under acidic conditions. The overexpression of PtOSH1 in A. thaliana promoted root development, fresh weight, and dry weight; upregulated the expression levels of ROS scavenging-related genes; and improved the activity of antioxidant enzymes, enhancing plant tolerance to cadmium (Cd) stress. This study aimed to provide guidance that will facilitate future studies of the function of PtOSH1 in the response of plants to Cd stress.

scholarly journals Identification and Characterization of Rice OsHKT1;3 Variants

Plants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2006
Author(s):  
Shahin Imran ◽  
Yoshiyuki Tsuchiya ◽  
Sen Thi Huong Tran ◽  
Maki Katsuhara
Keyword(s):  
Expression System ◽  
Ion Homeostasis ◽  
Transcript Level ◽  
Absolute Quantification ◽  
Xenopus Laevis Oocytes ◽  
Japonica Rice ◽  
Na Currents ◽  
Mrna Variants ◽  
Identification And Characterization

In rice, the high-affinity K+ transporter, OsHKT1;3, functions as a Na+-selective transporter. mRNA variants of OsHKT1;3 have been reported previously, but their functions remain unknown. In this study, five OsHKT1;3 variants (V1-V5) were identified from japonica rice (Nipponbare) in addition to OsHKT1;3_FL. Absolute quantification qPCR analyses revealed that the transcript level of OsHKT1;3_FL was significantly higher than other variants in both the roots and shoots. Expression levels of OsHKT1;3_FL, and some variants, increased after 24 h of salt stress. Two electrode voltage clamp experiments in a heterologous expression system using Xenopus laevis oocytes revealed that oocytes expressing OsHKT1;3_FL and all of its variants exhibited smaller Na+ currents. The presented data, together with previous data, provide insights to understanding how OsHKT family members are involved in the mechanisms of ion homeostasis and salt tolerance in rice.

scholarly journals Two bacterial glycosphingolipid synthases responsible for the synthesis of glucuronosylceramide and α-galactosylceramide

2020 ◽  
Vol 295 (31) ◽  
pp. 10709-10725
Author(s):  
Nozomu Okino ◽  
Mengbai Li ◽  
Qingjun Qu ◽  
Tomoko Nakagawa ◽  
Yasuhiro Hayashi ◽  
...  
Keyword(s):  
Gel Filtration ◽  
Homologous Gene ◽  
Bacterial Physiology ◽  
E Coli ◽  
Donor And Acceptor ◽  
Low Levels ◽  
Killer T Cells ◽  
Invariant Natural Killer ◽  
Identification And Characterization

Bacterial glycosphingolipids such as glucuronosylceramide and galactosylceramide have been identified as ligands for invariant natural killer T cells and play important roles in host defense. However, the glycosphingolipid synthases required for production of these ceramides have not been well-characterized. Here, we report the identification and characterization of glucuronosylceramide synthase (ceramide UDP-glucuronosyltransferase [Cer-GlcAT]) in Zymomonas mobilis, a Gram-negative bacterium whose cellular membranes contain glucuronosylceramide. On comparing the gene sequences that encode the diacylglycerol GlcAT in bacteria and plants, we found a homologous gene that is widely distributed in the order Sphingomonadales in the Z. mobilis genome. We first cloned the gene and expressed it in Escherichia coli, followed by protein purification using nickel–Sepharose affinity and gel filtration chromatography. Using the highly enriched enzyme, we observed that it has high glycosyltransferase activity with UDP-glucuronic acid and ceramide as sugar donor and acceptor substrate, respectively. Cer-GlcAT deletion resulted in a loss of glucuronosylceramide and increased the levels of ceramide phosphoglycerol, which was expressed in WT cells only at very low levels. Furthermore, we found sequences homologous to Cer-GlcAT in Sphingobium yanoikuyae and Bacteroides fragilis, which have been reported to produce glucuronosylceramide and α-galactosylceramide, respectively. We expressed the two homologs of the cer-glcat gene in E. coli and found that each gene encodes Cer-GlcAT and Cer-galactosyltransferase, respectively. These results contribute to the understanding of the roles of bacterial glycosphingolipids in host–bacteria interactions and the function of bacterial glycosphingolipids in bacterial physiology.

Genome-Wide Identification and Characterization of the FAR1/FHY3 Family in Populus trichocarpa Torr. & Gray and Expression Analysis in Light Response

Forests ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1385
Author(s):  
Jiujun Du ◽  
Lei Zhang ◽  
Xiaolan Ge ◽  
Xiaodong Xiang ◽  
Demei Cao ◽  
...  
Keyword(s):  
Regulatory Networks ◽  
Higher Plants ◽  
Populus Trichocarpa ◽  
Expression Patterns ◽  
Light Response ◽  
Cis Acting ◽  
Poplar Genome ◽  
Genome Wide ◽  
Identification And Characterization

Light is an important environmental factor for plant growth, and in higher plants, phytochrome A (phyA) is the predominant far-red photoreceptor, involved in various photoresponses. The FAR1/FHY3 transcription factor family, derived from transposases, is able to regulate plant development in response to multiple photosensitizers phytochrome. In total, 51 PtrFRSs were identified in the poplar genome, and were divided into 4 subfamilies. Among them, 47 PtrFRSs are located on 17 chromosomes. Upstream cis-acting elements of the PtrFRS genes were classified into three categories: growth and metabolism, stress and hormone, and the hormone and stress categories contained most of the cis-acting elements. Analysis of the regulatory networks and expression patterns showed that most PtrFRSs responded to changes in light intensity and were involved in the regulation of phytochromes. In this study, 51 PtrFRSs were identified and comprehensively bioinformatically analyzed, and preliminary functional analysis and prediction of PtrFRSs was carried out.

scholarly journals Identification and characterization of the Populus trichocarpa CLE family

BMC Genomics ◽  
2016 ◽  
Vol 17 (1) ◽  
Author(s):  
Huibin Han ◽  
Guohua Zhang ◽  
Mengyao Wu ◽  
Guodong Wang
Keyword(s):  
Populus Trichocarpa ◽  
Identification And Characterization

scholarly journals In Silico Identification and Characterization of N-Terminal Acetyltransferase Genes of Poplar (Populus trichocarpa)

2014 ◽  
Vol 15 (2) ◽  
pp. 1852-1864 ◽  
Author(s):  
Hang-Yong Zhu ◽  
Chun-Ming Li ◽  
Li-Feng Wang ◽  
Hui Bai ◽  
Yan-Ping Li ◽  
...  
Keyword(s):  
In Silico ◽  
Populus Trichocarpa ◽  
In Silico Identification ◽  
Identification And Characterization
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