scholarly journals Functional characterization of a DNA-dependent AAA ATPase in SimranZ1, a F cluster mycobacteriophage

2021 ◽  
Author(s):  
Ritam Das ◽  
Urmi Bajpai
Keyword(s):  
Dna Repair ◽  
Therapeutic Potential ◽  
Specific Binding ◽  
Functional Characterization ◽  
Microtiter Plate ◽  
Microtiter Plate Assay ◽  
Aaa Atpase ◽  
Maintenance Mechanism ◽  
Transmission Electron

AbstractMycobacteriophages are viruses of Mycobacterium spp. with promising diagnostic and therapeutic potential. Phage genome exploration and characterization of their proteomes are essential to gain a better understanding of their role in phage biology. So far, about 2014 mycobacteriophages have been genomically defined and 1563 phage protein families (phamilies) are identified. However, the function of only a fraction (about 15%) is known and a majority of ORFs in phage genomes are hypothetical proteins. In this study, from the annotated genome of a F1 cluster mycobacteriophage SimranZ1, a putative AAA ATPase (Gp65, Pham 9410) is characterized as a DNA-dependent ATPase. Sequence-based functional annotation predicted Gp65 to belong to the P-loop NTPase superfamily, having AAA_24 and RecA/RadA domains which are known to be involved in ATP-dependent DNA repair/maintenance mechanism. On molecular docking, Gly21 and Ser23 of Gp65 showed specific binding with ATP. Using a microtiter plate assay, ATPase activity of Gp65 was experimentally verified which was found to increase in the presence of dsDNA. Gel electrophoresis under non-denaturing condition showed the oligomeric states of Gp65 and Transmission Electron Microscopy revealed it to exist as a hexamer having a prominent central pore with a diameter of 1.9 nm. In summary, functional characterization of Gp65 as a DNA dependent AAA ATPase indicates its role in DNA repair/maintenance mechanism in mycobacteriophages.

Functional characterization of the Caenorhabditis elegans DNA repair enzyme APN-1

DNA Repair ◽  
2012 ◽  
Vol 11 (10) ◽  
pp. 811-822 ◽  
Author(s):  
Xiaoming Yang ◽  
Jinjiang Fan ◽  
Alexander A. Ishchenko ◽  
Devang Patel ◽  
Murat K. Saparbaev ◽  
...  
Keyword(s):  
Dna Repair ◽  
Caenorhabditis Elegans ◽  
Functional Characterization ◽  
Repair Enzyme

CHARACTERIZATION OF BACTERIAL ADHESION AND BIOFILM FORMATION ABILITY OF LIVESTOCK-ASSOCIATED METHICILLIN-RESISTANTSTAPHYLOCOCCUS AUREUSISOLATED FROM SWINE AND SLAUGHTERHOUSE WASTEWATERS

2014 ◽  
Vol 40 (02) ◽  
pp. 101-107
Author(s):  
Min-Tao Wan ◽  
Chin-Cheng Chou
Keyword(s):  
Bacterial Adhesion ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Ecological Niches ◽  
Microtiter Plate Assay ◽  
Environmental Group ◽  
Biofilm Production ◽  
The Difference ◽  
Wastewater Samples

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST9 has emerged as a potential zoonotic pathogen for humans and animals. Bacterial adhesion factors and biofilms mediate host colonization and infection of MRSA. This study investigated the dynamics of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), biofilm formation gene (intercellular adhesion [ica]), and biofilm expression in MRSA from the nasal samples of asymptomatic pigs (the nasal group, n = 147) and swine slaughterhouse wastewater samples (the environmental group, n = 86). Biofilm formation was quantified by microtiter plate assay. The most prevalent MSCRAMM profile was clfA-clfB-spa-eno-ebps-fib and more than 70% of the LA-MRSA ST9 isolates harbored the biofilm formation gene. Environmental MRSA harbored lower levels of the ica locus and MSCRAMMs (clfA and fib) than did the nasal group, suggesting possible gene loss. Biofilm production in the nasal group was higher than in the environmental group, indicating the difference in biofilm formation in MRSA isolates from different ecological niches. The higher prevalence of MSCRAMMs, biofilm formation gene, and biofilm production in LA-MRSA ST9 may enhance the persistence and infectivity of MRSA in the swine population and present a threat to the health of livestock as well as farm workers.

scholarly journals Functional characterization of polymorphisms in DNA repair genes using cytogenetic challenge assays.

2003 ◽  
Vol 111 (15) ◽  
pp. 1843-1850 ◽  
Author(s):  
William W Au ◽  
Salama A Salama ◽  
Carlos H Sierra-Torres
Keyword(s):  
Dna Repair ◽  
Functional Characterization ◽  
Dna Repair Genes ◽  
Repair Genes

scholarly journals Characterization of Biofilm production in XDR Proteus mirabilis isolated from Baghdad city hospitals

2020 ◽  
Vol 11 (2) ◽  
pp. 70-78
Author(s):  
Rawa Aziz ◽  
◽  
Elaf Mohameed
Keyword(s):  
Electron Microscope ◽  
Antibiotic Sensitivity ◽  
Microtiter Plate ◽  
Automated System ◽  
Drug Resistant ◽  
Resistance Level ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Biofilm Production

Objective(s): P. mirabilis is considered as extensive drug resistant pathogen in many studies as it can resist complex antibiotic regimes, such pathogen can be threat to public health, especially when it has the ability to produce biofilm. Therefore, biofilm production was characterized among XDR P. mirabilis local isolates in this research. Material and Methods: 100 P. mirabilis isolated from wound infections from patients admitted to Baghdad hospitals. They were identified using biochemical test and Vitek2 system. The MIC test for antibiotic sensitivity was done by Vitek 2 automated system. Biofilm production was identified phenotypically by twitching motility test, scanning electron microscope, and microtiter plate assay. Results: It could be revealed that 8/100 isolates were MDR, 90/100 isolates were XDR and pan drug resistance level was shown in only two isolates. 80% of isolates had motile ability through twitching assay, and scan electron microscope study revealed that 76% of XDR isolates could produce different stages of biofilm on coverslip placed in MacConkey broth. Microtiter plate assay revealed 81% of XDR isolates were biofilm producers. Conclusion: it could be concluded that extensive drug resistant P. mirabilis can produce biofilm hence resist several important antibiotics; making treatment of infection among wounded patients is such a challenge in many hospitals in Baghdad.

scholarly journals Yeast-based assays for the functional characterization of cancer-associated variants of human DNA repair genes

2020 ◽  
Vol 7 (7) ◽  
pp. 162-174
Author(s):  
Tiziana Cervelli ◽  
Samuele Lodovichi ◽  
Francesca Bellè ◽  
Alvaro Galli
Keyword(s):  
Dna Repair ◽  
Functional Characterization ◽  
Dna Repair Genes ◽  
Human Dna ◽  
Repair Genes

O IX.2 Molecular cloning and functional characterization of the human XRCC2 DNA repair gene

1997 ◽  
Vol 379 (1) ◽  
pp. S61
Author(s):  
John Thacker ◽  
Richard Cartwright ◽  
Cathryn Tambini ◽  
Andrew George ◽  
Johanna Rommens ◽  
...  
Keyword(s):  
Dna Repair ◽  
Molecular Cloning ◽  
Functional Characterization ◽  
Repair Gene ◽  
Dna Repair Gene
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