Gp41, a superfamily SF2 helicase from bacteriophage BFK20

Halgasova, Nora; Matuskova, Radka; Kraus, Daniel; Tkacova, Adela; Balusikova, Lenka; Bukovská, Gabriela

doi:10.1016/j.virusres.2017.12.005

Cited by 4 publications

(2 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover, reactions are rapid and sensitive. Therefore, the opportunity of detecting and computing bacteria species through specifi c enzymatic functional model activities has been under study for many years now [30][31][32].…”

Section: The Assembly Of Bacteriophage Functional Enzymatic Models Inmentioning

confidence: 99%

The Assembly of Bacteriophage Functional Enzymatic Models in Association with E. coli Proteins’ Profiles

Elshayeb¹,

Elfatih²,

Salih³

et al. 2020

J Biomed Res Environ Sci

View full text Add to dashboard Cite

Introduction: The invasion of bacteriophage on the associated host bacterium depends on their receptors' orientation that adsorb them to cell surface. During phage replication a valuable number of proteins acts as lytic enzymes for host puncher at the beginning of the infection and other for burst after lytic cycle compilation. Accordingly, the proteomic relationship among phage and bacterium proteins could easily be studied by their protein profiles analysis. Objective: To detect bacteriophages functional enzymes during lytic cycle. Methods: The isolation and identification of Escherichia coli and their parasitic T7 phage group was done using bacterial culture and common plaque assay techniques. The investigations and protein-protein interactions' assays were inveterate by proteins profile of phage and bacterium using Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) to find out their molecular weights, where the scaled location of each mobile band was compared to the standards of identified proteins weights in the molecular ladder. Thereafter, Protein model's assembly and bands migration was done by computer analytical software. Results: Mobilization of the phage' proteins inside the Two Dimensions (2D) gel ranged between 60 and 12 kDa where a model of 4 main bands with molecular weights of (46, 35, 24 and 14 kDa) is corresponded to the host ones, where pure 9 bands with molecular weight ranged between 96-24 kDa. The computational model analysis showed common shared molecular masses of 47, 34 and 16 kDa on plot area of the phage and the bacterium. Model interpretation confirmed that proteins ranged from 47.7 to 34.3 kDa resembles 43.3% of whole phage's proteins that assembled the capsid head and the coil, while the molecular weight mass of 22.5 formed the tail's proteins. The lytic enzymes' molecular weight was ranged between 18-14 kDa according to the function of the enzyme. The study revealed that the 34 kDa band has the common shared peak between T7 phage group and associated Escherichia coli host. Conclusion: Functional models of analysed proteins during phage assembly, ensures lytic enzymes are built in the capsid head and the lysozyme in the tail, they facilitate the enzymatic decay for bacterial host. This enzymatic function is related to the lytic cycle of the bacteriophages and their phenomenon in employing the bacterial DNA in proteins manufacturing during their replication inside host.

show abstract

Section: The Assembly Of Bacteriophage Functional Enzymatic Models Inmentioning

confidence: 99%

The Assembly of Bacteriophage Functional Enzymatic Models in Association with E. coli Proteins’ Profiles

Elshayeb¹,

Elfatih²,

Salih³

et al. 2020

J Biomed Res Environ Sci

View full text Add to dashboard Cite

show abstract

“…Helicases, i.e. enzymes that catalyze the unfolding of DNA or RNA, are often produced by phage genomes 75 . Most notably, DEAD/DEAH-box containing helicases participate in RNA metabolism in many, essential ways, such as by regulating gene expression and signalling 76 .…”

Section: Other Predicted Proteinsmentioning

confidence: 99%

Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Kyrkou

Carstens

Ellegaard-Jensen

et al. 2019

Preprint

View full text Add to dashboard Cite

Bacteria of Lactobacillus sp. are very useful to humans. However, the biology and genomic diversity of their (bacterio)phage enemies remains understudied. Knowledge on Lactobacillus phage diversity should broaden to develop efficient phage control strategies. To this end, organic waste samples were screened for phages against two wine-related Lactobacillus plantarum strains. Isolates were shotgun sequenced and compared against the phage database and each other by phylogenetics and comparative genomics. The new isolates had only three distant relatives from the database but displayed a high overall degree of genomic similarity amongst them. The latter allowed for the use of one isolate as a representative to conduct transmission electron microscopy and structural protein sequencing, and to study phage adsorption and growth kinetics. The microscopy and proteomics tests confirmed the observed diversity of the new isolates and supported their classification to the family Siphoviridae and the proposal of the new phage genus "Silenusvirus". Methods Environmental samples, phage assays and bacterial strainsLactobacillus phages were isolated from organic household waste samples. The samples were collected from two different organic waste treatment plants in Denmark (treatment plants A and B). Initially, the samples were split into two subsamples and processed as detailed in Kyrkou et al 12 . The resulting filtrates were screened for phages using the double agar overlay method 16 and a top layer of 0.4% w/v agarose. Specifically, efficiency of plating assays were performed against two indicator strains that had earlier been acquired from private collections, L. plantarum L1 (wine fermentation isolate) and L. plantarum MW-1 (grape isolate). Single plaques were resuspended in 0.7 mL SM buffer and later filtered by 0.45-µm pore size PVDF spin filters (Ciro, Florida, USA). Each purified plaque underwent two further reisolation-filtration cycles to ensure pure stock cultures and was stored at 4°C. For transmission electron microscopy (TEM) and protein sequencing, lysates of 10 10 plaque-forming units (PFUs)/mL) were further purified and concentrated using caesium chloride (CsCl) gradient ultracentrifugation, as described elsewhere 17 . All incubations of phage manipulations were done at 25°C using De Man, Rogosa and Sharpe (MRS) broth and agar media supplemented with 10 mM CaCl 2 (MRSΦ), whereas indicator strains were grown in MRS at 37°C. TEM analysis and structural protein sequencingPhage morphology and structural proteins were determined using the CsCl-purified stocks. Micrographs of phage Silenus were generated as in other studies 12,18 . The mean values and standard deviations of all Silenus virion dimensions were elucidated after inspection of 20-23 phage particles. Structural proteins were sequenced following published protocols 12,18 . Briefly, 100 µL of the CsCl-purified stocks were filtered through an Amicon Ultra filter unit (MWCO 30k Da) and desalted four times. Phage particles (10 µL) were denaturised in 6 M urea, ...

show abstract

Characterization of a newly discovered putative DNA replication initiator from Paenibacillus polymyxa phage phiBP

Halgasova

Javorova

Bocanova

et al. 2023

Microbiological Research

View full text Add to dashboard Cite

Gp41, a superfamily SF2 helicase from bacteriophage BFK20

Cited by 4 publications

References 35 publications

The Assembly of Bacteriophage Functional Enzymatic Models in Association with E. coli Proteins’ Profiles

The Assembly of Bacteriophage Functional Enzymatic Models in Association with E. coli Proteins’ Profiles

Isolation and characterisation of novel phages infectingLactobacillus plantarumand proposal of a new genus, “Silenusvirus”

Characterization of a newly discovered putative DNA replication initiator from Paenibacillus polymyxa phage phiBP

Contact Info

Product

Resources

About