scholarly journals Phosphoglycerate kinase: structural aspects and functions, with special emphasis on the enzyme from Kinetoplastea

Open Biology ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 200302
Author(s):  
Maura Rojas-Pirela ◽  
Diego Andrade-Alviárez ◽  
Verónica Rojas ◽  
Ulrike Kemmerling ◽  
Ana J. Cáceres ◽  
...  
Keyword(s):  
Phosphoglycerate Kinase ◽  
Glycolytic Enzyme ◽  
Open Reading Frames ◽  
Activity Regulation ◽  
Free Living ◽  
Structural Domains ◽  
Domains Of Life ◽  
Moonlighting Functions ◽  
Reading Frames ◽  
Structural Aspects

Phosphoglycerate kinase (PGK) is a glycolytic enzyme that is well conserved among the three domains of life. PGK is usually a monomeric enzyme of about 45 kDa that catalyses one of the two ATP-producing reactions in the glycolytic pathway, through the conversion of 1,3-bisphosphoglycerate (1,3BPGA) to 3-phosphoglycerate (3PGA). It also participates in gluconeogenesis, catalysing the opposite reaction to produce 1,3BPGA and ADP. Like most other glycolytic enzymes, PGK has also been catalogued as a moonlighting protein, due to its involvement in different functions not associated with energy metabolism, which include pathogenesis, interaction with nucleic acids, tumorigenesis progression, cell death and viral replication. In this review, we have highlighted the overall aspects of this enzyme, such as its structure, reaction kinetics, activity regulation and possible moonlighting functions in different protistan organisms, especially both free-living and parasitic Kinetoplastea. Our analysis of the genomes of different kinetoplastids revealed the presence of open-reading frames (ORFs) for multiple PGK isoforms in several species. Some of these ORFs code for unusually large PGKs. The products appear to contain additional structural domains fused to the PGK domain. A striking aspect is that some of these PGK isoforms are predicted to be catalytically inactive enzymes or ‘dead’ enzymes. The roles of PGKs in kinetoplastid parasites are analysed, and the apparent significance of the PGK gene duplication that gave rise to the different isoforms and their expression in Trypanosoma cruzi is discussed.

scholarly journals Vibrio natriegens, a new genomic powerhouse

2016 ◽  
Author(s):  
Henry H. Lee ◽  
Nili Ostrov ◽  
Brandon G. Wong ◽  
Michaela A. Gold ◽  
Ahmad S. Khalil ◽  
...  
Keyword(s):  
Recombinant Dna ◽  
Open Reading Frames ◽  
Host Organism ◽  
Recombinant Dna Technology ◽  
Free Living ◽  
Growth Properties ◽  
Culturing Conditions ◽  
Laboratory Culturing ◽  
Tools And Techniques ◽  
Reading Frames

Recombinant DNA technology has revolutionized biomedical research with continual innovations advancing the speed and throughput of molecular biology. Nearly all these tools, however, are reliant onEscherichia colias a host organism, and its lengthy growth rate increasingly dominates experimental time. Here we report the development ofVibrio natriegens, a free-living bacteria with the fastest generation time known, into a genetically tractable host organism. We systematically characterize its growth properties to establish basic laboratory culturing conditions. We provide the first completeVibrio natriegensgenome, consisting of two chromosomes of 3,248,023 bp and 1,927,310 bp that together encode 4,578 open reading frames. We reveal genetic tools and techniques for working withVibrio natriegens. These foundational resources will usher in an era of advanced genomics to accelerate biological, biotechnological, and medical discoveries.

scholarly journals Prediction of cellulolytic and hemicellulolytic bacterial diversity in the gut of Coptotermes gestroi in the Southern Vietnam

2020 ◽  
Vol 17 (3) ◽  
pp. 537-544
Author(s):  
Nguyen Thi Thao ◽  
Do Thi Huyen ◽  
Truong Nam Hai
Keyword(s):  
Bacterial Species ◽  
Abundant Species ◽  
Open Reading Frames ◽  
Metagenomic Dna ◽  
Free Living ◽  
Southern Vietnam ◽  
Degrading Bacteria ◽  
Coptotermes Gestroi ◽  
Reading Frames ◽  
Living Bacteria

In lower termite such as Coptotermes gestroi, cellulose and hemicellulose are hydrolysed by cellulases and hemicellulases secreted from bacteria, archaea, protozoa and fungy in the hindgut. In which, majority of the enzymes are contributed by protozoa. From the metagenomic DNA data (125,423 open reading frames -ORFs) of free-living bacteria in the gut of C. gestroi harvested in Southern Vietnam and by MEGA 4.0 software, 100.340 ORFs were classified into 1,368 species, 628 genera, 217 families, 97 orders, 41 classes and 22 phyla (Do et al., 2014). Among these, 2,131 ORFs (2,12%) belong to 24 bacterial species (account 1,75% bacterial species), 11 families, 9 orders, 8 classes and 5 phyla were predicted have ability to produce cellulases; 679 ORFs belong to 18 bacterial species 8 families, 6 orders, 5 classes, 4 phyla were predicted have ability to produce hemicellulase. Majority of cellulase producers were species which of Firmicutes (15/24 species), accumulated in class Clostridia, order Clostridiales. The most abundant cellulase producer was Pseudomonas fluorescens (1,258 ORFs) of order Pseudomonadaceae. Out of the 18 hemicellulase producers, the most abundant species was Clostridium thermocellum (113 ORFs) in the phylum Firmicutes, followed by 3 species belonging to the phylum Bacteroidetes. The species predicted to produce both cellulase, hemicellulase were C. thermocellum, Ruminococcusns flavefaciens and Bacillus subtilis. Our study provides  a data of gut cellulose and hemicellulose - degrading bacteria composition of C. gestroi

scholarly journals Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens

2021 ◽  
Vol 9 (5) ◽  
pp. 1005
Author(s):  
Olga Chervyakova ◽  
Elmira Tailakova ◽  
Nurlan Kozhabergenov ◽  
Sandugash Sadikaliyeva ◽  
Kulyaisan Sultankulova ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Viral Vector ◽  
Foot And Mouth Disease ◽  
Open Reading Frames ◽  
Foreign Gene ◽  
Mouth Disease Virus ◽  
Foreign Genes ◽  
Green Fluorescent ◽  
Reading Frames

Capripoxviruses with a host range limited to ruminants have the great potential to be used as vaccine vectors. The aim of this work was to evaluate attenuated sheep pox virus (SPPV) vaccine strain NISKHI as a vector expressing several genes. Open reading frames SPPV020 (ribonucleotide kinase) and SPPV066 (thymidine kinase) were selected as sites for the insertion of foreign genes. Two integration plasmids with expression cassette were designed and constructed. Recombinant SPPVs expressing an enhanced green fluorescent protein (EGFP) (rSPPV(RRΔ)EGFP and rSPPV(TKΔ)EGFP), Foot-and-mouth disease virus capsid protein (VP1), and Brucella spp. outer membrane protein 25 (OMP25) (rSPPV(RRΔ)VP1A-(TKΔ)OMP25) were generated under the transient dominant selection method. The insertion of foreign genes into the SPPV020 and SPPV066 open reading frames did not influence the replication of the recombinant viruses in the cells. Successful foreign gene expression in vitro was assessed by luminescent microscopy (EGFP) and Western blot (VP1 and OMP25). Our results have shown that foreign genes were expressed by rSPPV both in permissive (lamb testicles) and non-permissive (bovine kidney, saiga kidney, porcine kidney) cells. Mice immunized with rSPPV(RRΔ)VP1A-(TKΔ)OMP25 elicited specific antibodies to both SPPV and foreign genes VP1 and OMP25. Thus, SPPV NISKHI may be used as a potential safe immunogenic viral vector for the development of polyvalent vaccines.

Sign in / Sign up

Export Citation Format

Share Document