DDX17 helicase promotes resolution of R-loop-mediated transcription-replication conflicts in human cells

R-loops are three-stranded nucleic acid structures composed of an RNA:DNA hybrid and displaced DNA strand. These structures can halt DNA replication when formed co-transcriptionally in the opposite orientation to replication fork progression. A recent study has shown that replication forks stalled by co-transcriptional R-loops can be restarted by a mechanism involving fork cleavage by MUS81 endonuclease, followed by ELL-dependent reactivation of transcription, and fork religation by the DNA ligase IV (LIG4)/XRCC4 complex. However, how R-loops are eliminated to allow the sequential restart of transcription and replication in this pathway remains elusive. Here, we identified the human DDX17 helicase as a factor that associates with R-loops and counteracts R-loop-mediated replication stress to preserve genome stability. We show that DDX17 unwinds R-loops in vitro and promotes MUS81-dependent restart of R-loop-stalled forks in human cells in a manner dependent on its helicase activity. Loss of DDX17 helicase induces accumulation of R-loops and the formation of R-loop-dependent anaphase bridges and micronuclei. These findings establish DDX17 as a component of the MUS81-LIG4-ELL pathway for resolution of R-loop-mediated transcription-replication conflicts, which may be involved in R-loop unwinding

B-VNF: Blockchain-enhanced Architecture for VNF Orchestration in MEC-5G Networks

ELECTR NETWORKThe roll-out of 5G technology will nurture the realization of broadband, ultra-reliable, and zero latency services. Network Function Virtualization (NFV) and Multi-Access Edge Computing (MEC) are among the key enablers for 5G. The synergy between NFV and MEC allows migration of Virtual Network Functions (VNF) from cloud to the edge of the network thereby adding agility to the softwarized 5G networks. The overall orchestration of VNF includes, but is not limited to, processing VNF requests, selecting appropriate VNF, migrating VNF from cloud to MEC, instantiating migrated VNF at MEC, settling payment according to a VNF's usage, maintaining VNF's reputation, etc. The orchestration is not foolproof and raises doubts about its trustworthiness. To address all the existing issues in a unified manner, we leverage Blockchain technology as yet another enabling technology for MEC-enabled 5G. Thus, we propose a Blockchain-enhanced architecture for secure VNF orchestration such that issues like authenticity, integrity, confidentiality, reputation, payment transfer, and many more are resolved. To furnish a Proof-of-Concept (PoC), we develop a prototypical DApp (Decentralized Application) using Ethereum Blockchain and Suricata as an exemplar VNF. Further, we discuss the strong resiliency of the proposed architecture against numerous well-known attacks.European Commission Horizon 2020Academy of Finland in 6Genesis FlagshipEuropean Union in RESPONSE 5

Mycobacterium tuberculosis UvrD1 and UvrD2 helicases unwind G-quadruplex DNA

Unresolved G-quadruplex (G4) DNA secondary structures impede DNA replication and can lead to DNA breaks and to genome instability. Helicases are known to unwind G4 structures and thereby facilitate genome duplication. Escherichia coli UvrD is a multifunctional helicase that participates in DNA repair, recombination and replication. Previously, we had demonstrated a novel role of E. coli UvrD helicase in resolving G4 structures. Mycobacterium tuberculosis genome encodes two orthologs of E. coli UvrD helicase, UvrD1 and UvrD2. It is unclear whether UvrD1 or UvrD2 or both helicases unwind G4 DNA structures. Here, we demonstrate that M. tuberculosis UvrD1 and UvrD2 unwind G4 tetraplexes. Both helicases were proficient in resolving previously characterized tetramolecular G4 structures in an ATP hydrolysis and single-stranded 3 `-tail-dependent manner. Notably, M. tuberculosis UvrD1 and UvrD2 were efficient in unwinding G4 structures derived from the potential G4 forming sequences present in the M. tuberculosis genome. These data suggest an extended role for M. tuberculosis UvrD1 and UvrD2 helicases in resolving G4 DNA structures and provide insights into the maintenance of genome integrity via G4 DNA resolution

Escherichia coli and Neisseria gonorrhoeae UvrD helicase unwinds G4 DNA structures

G-quadruplex (G4) secondary structures have been implicated in various biological processes, including gene expression, DNA replication and telomere maintenance. However, unresolved G4 structures impede replication progression which can lead to the generation of DNA double-strand breaks and genome instability. Helicases have been shown to resolve G4 structures to facilitate faithful duplication of the genome. Escherichia coli UvrD (EcUvrD) helicase plays a crucial role in nucleotide excision repair, mismatch repair and in the regulation of homologous recombination. Here, we demonstrate a novel role of E. coli and Neisseria gonorrhoeae UvrD in resolving G4 tetraplexes. EcUvrD and N. gonorrhoeae UvrD were proficient in unwinding previously characterized tetramolecular G4 structures. Notably, EcUvrD was equally efficient in resolving tetramolecular and bimolecular G4 DNA that were derived from the potential G4-forming sequences from the genome of E. coli. Interestingly, in addition to resolving intermolecular G4 structures, EcUvrD was robust in unwinding intramolecular G4 structures. These data for the first time provide evidence for the role of UvrD in the resolution of G4 structures, which has implications for the in vivo role of UvrD helicase in G4 DNA resolution and genome maintenance

B-VNF:blockchain-enhanced architecture for VNF orchestration in MEC-5G networks

Abstract The roll-out of 5G technology will nurture the realization of broadband, ultra-reliable, and zero latency services. Network Function Virtualization (NFV) and Multi-Access Edge Computing (MEC) are among the key enablers for 5G. The synergy between NFV and MEC allows migration of Virtual Network Functions (VNF) from cloud to the edge of the network thereby adding agility to the softwarized 5G networks. The overall orchestration of VNF includes, but is not limited to, processing VNF requests, selecting appropriate VNF, migrating VNF from cloud to MEC, instantiating migrated VNF at MEC, settling payment according to a VNF’s usage, maintaining VNF’s reputation, etc. The orchestration is not foolproof and raises doubts about its trustworthiness. To address all the existing issues in a unified manner, we leverage Blockchain technology as yet another enabling technology for MEC-enabled 5G. Thus, we propose a Blockchain-enhanced architecture for secure VNF orchestration such that issues like authenticity, integrity, confidentiality, reputation, payment transfer, and many more are resolved. To furnish a Proof-of-Concept (PoC), we develop a prototypical DApp (Decentralized Application) using Ethereum Blockchain and Suricata as an exemplar VNF. Further, we discuss the strong resiliency of the proposed architecture against numerous well-known attacks

Sialoglycosylation of RBC in Visceral Leishmaniasis Leads to Enhanced Oxidative Stress, Calpain-Induced Fragmentation of Spectrin and Hemolysis

Visceral leishmaniasis (VL) caused by the intracellular parasite Leishmania donovani accounts for an estimated 12 million cases of human infection. It is almost always associated with anemia, which severely complicates the disease course. However, the pathological processes leading to anemia in VL have thus far not been adequately characterized to date. In studying the glycosylation patterns of peripheral blood cells we found that the red blood cells (RBC) of VL patients (RBCVL) express eight 9-O-acetylated sialoglycoproteins (9-O-AcSGPs) that are not detected in the RBC of healthy individuals (RBCN). At the same time, the patients had high titers of anti-9-O-AcSGP IgG antibodies in their sera. These two conditions appear to be linked and related to the anemic state of the patients, as exposure of RBCVL but not RBCN to anti-9-O-AcSGPs antibodies purified from patient sera triggered a series of responses. These included calcium influx via the P/Q-type but not L-type channels, activation of calpain I, proteolysis of spectrin, enhanced oxidative stress, lipid peroxidation, externalization of phosphatidyl serine with enhanced erythrophagocytosis, enhanced membrane fragility and, finally, hemolysis. Taken together, this study suggests that the enhanced hemolysis is linked to an impairment of membrane integrity in RBCVL which is mediated by ligand-specific interaction of surface 9-O-AcSGPs. This affords a potential explanation for the structural and functional features of RBCVL which are involved in the hemolysis related to the anemia which develops in VL patients

Abstracts of National Conference on Research and Developments in Material Processing, Modelling and Characterization 2020

This book presents the abstracts of the papers presented to the Online National Conference on Research and Developments in Material Processing, Modelling and Characterization 2020 (RDMPMC-2020) held on 26th and 27th August 2020 organized by the Department of Metallurgical and Materials Science in Association with the Department of Production and Industrial Engineering, National Institute of Technology Jamshedpur, Jharkhand, India. Conference Title: National Conference on Research and Developments in Material Processing, Modelling and Characterization 2020Conference Acronym: RDMPMC-2020Conference Date: 26–27 August 2020Conference Location: Online (Virtual Mode)Conference Organizer: Department of Metallurgical and Materials Engineering, National Institute of Technology JamshedpurCo-organizer: Department of Production and Industrial Engineering, National Institute of Technology Jamshedpur, Jharkhand, IndiaConference Sponsor: TEQIP-