Base excision repair initiated rolling circle amplification-based fluorescent assay for screening uracil-DNA glycosylase activity using Endo IV-assisted cleavage of AP probes

The Analyst ◽  
2018 ◽  
Vol 143 (16) ◽  
pp. 3951-3958 ◽  
Author(s):  
Jingfeng Wang ◽  
Yu Wang ◽  
Su Liu ◽  
Haiwang Wang ◽  
Xue Zhang ◽  
...  
Keyword(s):  
Base Excision Repair ◽  
Signal Amplification ◽  
Excision Repair ◽  
Rolling Circle Amplification ◽  
Cost Effective ◽  
Ultrasensitive Detection ◽  
Fluorescent Assay ◽  
Uracil Dna Glycosylase ◽  
Rolling Circle ◽  
Base Excision

A simple, robust and cost effective biosensing platform for the ultrasensitive detection of UDG activity was established based on base excision repair-initiated primer generation for RCA with Endo IV-assisted signal amplification.

scholarly journals Uracil-DNA Glycosylase in Base Excision Repair and Adaptive Immunity

2011 ◽  
Vol 286 (19) ◽  
pp. 16669-16680 ◽  
Author(s):  
Berit Doseth ◽  
Torkild Visnes ◽  
Anders Wallenius ◽  
Ida Ericsson ◽  
Antonio Sarno ◽  
...  
Keyword(s):  
Adaptive Immunity ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Dna Glycosylase ◽  
Uracil Dna Glycosylase ◽  
Base Excision

scholarly journals The p53-Induced Oncogenic Phosphatase PPM1D Interacts with Uracil DNA Glycosylase and Suppresses Base Excision Repair

2004 ◽  
Vol 15 (4) ◽  
pp. 621-634 ◽  
Author(s):  
Xiongbin Lu ◽  
Dora Bocangel ◽  
Bonnie Nannenga ◽  
Hiroshi Yamaguchi ◽  
Ettore Appella ◽  
...  
Keyword(s):  
Base Excision Repair ◽  
Excision Repair ◽  
Dna Glycosylase ◽  
Uracil Dna Glycosylase ◽  
Base Excision

scholarly journals Base-Excision Repair Activity of Uracil-DNA Glycosylase Monitored Using the Latch Zone of α-Hemolysin

2013 ◽  
Vol 135 (51) ◽  
pp. 19347-19353 ◽  
Author(s):  
Qian Jin ◽  
Aaron M. Fleming ◽  
Robert P. Johnson ◽  
Yun Ding ◽  
Cynthia J. Burrows ◽  
...  
Keyword(s):  
Base Excision Repair ◽  
Excision Repair ◽  
Dna Glycosylase ◽  
Uracil Dna Glycosylase ◽  
Repair Activity ◽  
Base Excision

scholarly journals When UDG and hAPE1 Meet Cyclopurines. How (5′R) and (5′S) 5′,8-Cyclo-2′-deoxyadenosine and 5′,8-Cyclo-2′-deoxyguanosine Affect UDG and hAPE1 Activity?

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5177
Author(s):  
Michał Szewczuk ◽  
Karolina Boguszewska ◽  
Julia Kaźmierczak-Barańska ◽  
Bolesław T. Karwowski
Keyword(s):  
Base Excision Repair ◽  
Excision Repair ◽  
Dna Lesions ◽  
Repair System ◽  
Cellular Mechanisms ◽  
Uracil Dna Glycosylase ◽  
Base Excision ◽  
Base Excision Repair System ◽  
Synthetic Oligonucleotides ◽  
Ap Site

Ionizing radiation is a factor that seriously damages cellular mechanisms/macromolecules, e.g., by inducing damage in the human genome, such as 5′,8-cyclo-2′-deoxypurines (cdPus). CdPus may become a component of clustered DNA lesions (CDL), which are notably unfavorable for the base excision repair system (BER). In this study, the influence of 5′S and 5′R diastereomers of 5′,8-cyclo-2′-deoxyadenosine (cdA) and 5′,8-cyclo-2′-deoxyguanosine (cdG) on the uracil-DNA glycosylase (UDG) and human AP site endonuclease 1 (hAPE1) activity has been taken under consideration. Synthetic oligonucleotides containing 2′-deoxyuridine (dU) and cdPu were used as a model of single-stranded CDL. The activity of the UDG and hAPE1 enzymes decreased in the presence of RcdG compared to ScdG. Contrary to the above, ScdA reduced enzyme activity more than RcdA. The presented results show the influence of cdPus lesions located within CDL on the activity of the initial stages of BER dependently on their position toward dU. Numerous studies have shown the biological importance of cdPus (e.g., as a risk of carcinogenesis). Due to that, it is important to understand how to recognize and eliminate this type of DNA damage from the genome.

scholarly journals Deficient Uracil Base Excision Repair Leads to Persistent dUMP in HIV Proviruses During Infection of Monocytes and Macrophages

2020 ◽  
Author(s):  
Mesfin Meshesha ◽  
Alexandre Esadze ◽  
Junru Cui ◽  
Natela Churgulia ◽  
Sushil Kumar Sahu ◽  
...  
Keyword(s):  
Base Excision Repair ◽  
Reverse Transcription ◽  
Excision Repair ◽  
Dna Glycosylase ◽  
Target Cells ◽  
Viral Dna ◽  
Uracil Dna Glycosylase ◽  
Monocytes And Macrophages ◽  
Base Excision ◽  
Uracil Base

Abstract Background Non-dividing cells of the myeloid lineage such as monocytes and macrophages are target cells of HIV that have low dNTP pool concentrations and elevated levels of dUTP, which leads to frequent incorporation of dUMP opposite to A during reverse transcription (“uracilation”). One factor determining the fate of dUMP in proviral DNA is the host cell uracil base excision repair (UBER) system. Here we explore the relative UBER capacity of monocytes (MC) and monocyte-derived macrophages (MDM) and the fate of integrated uracilated viruses in both cell types to understand the implications of viral dUMP on HIV diversification and infectivity.Results We find that monocytes are almost completely devoid of functional UBER, while macrophages are mainly deficient in the initial enzyme uracil DNA glycosylase (hUNG2). Accordingly, dUMP persists in viral DNA during the lifetime of a MC and can only be removed after differentiation of MC into MDM. Overexpression of human uracil DNA glycosylase in MDM prior to infection resulted in rapid removal of dUMP from HIV cDNA and near complete depletion of dUMP-containing viral copies. This finding establishes that the low hUNG2 expression level in these cells limits UBER but that hUNG2 is restrictive against uracilated viruses. In contrast, overexpression of hUNG2 after viral integration did not accelerate the excision of uracils, suggesting that they may poorly accessible in the context of chromatin. We found that viral DNA molecules with incorporated dUMP contained unique (+) strand transversion mutations that were not observed when dUMP was absent (G→T, T→A, T→G, A→C). These observations and other considerations suggest that dUMP introduces errors predominantly during (-) strand synthesis when the template is RNA. These mutations may arise from the increased mispairing and duplex destabilizing effects of dUMP relative to dTMP during reverse transcription. Overall, the likelihood of producing a functional virus from in vitro infection of MC is about 50-fold and 300-fold reduced as compared to MDM and activated T cells.Conclusions The results implicate viral dUMP incorporation in MC and MDM as a potential viral diversification and restriction pathway during human HIV infection.

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