Query: RPA

title: The Future Digital Work Force: Robotic Process Automation (RPA)
abstract: The Robotic Process Automation (RPA) is a new wave of future technologies. Robotic Process Automation is one of the most advanced technologies in the area of computers science, electronic and communications, mechanical engineering, and information technology. It is a combination of both hardware and software, networking and automation for doing things very simple. In this light, the research manuscript investigated the secondary data which is available on google, academic and research databases. The investigation went for totally 6 months, i.e. 1 1 2018 to 30 6 2018. A very few empirical articles, white papers, blogs and were found RPA and came across to compose this research manuscript. This study is exploratory in nature because of the contemporary phenomenon. The keywords used in searching of the database were Robotic Process Automation, RPA, Robots, Artificial Intelligence, Blue Prism. The study finally discovered that Robots and Robotic Process Automation technologies are becoming compulsory as a part to do business operations in organizations across the globe. Robotic Process Automation can bring immediate value to the core business processes including employee payroll, employee status changes, new hire recruitment, and onboarding, accounts receivable and payable, invoice processing, inventory management, report creation, software installations, data migration, and vendor onboarding etc. to name a few applications. Besides, the Robotic Process Automation has abundant applications including healthcare and pharmaceuticals, financial services, outsourcing, retail, telecom, energy and utilities, real estate and FMCG and many more sectors. To put in the right place of RPA in business operations, their many allied technologies are working at the background level, artificial intelligence, machine learning, deep learning, data analytics, HR analytics, virtual reality (second life) home automation, blockchain technologies, 4D printing etc. Moreover, it covers the content of different start ups companies and existing companies their RPA applications used across the world. This manuscript will be a good guideline for the academicians, researchers, students, and practitioners to get an overall idea.

title: Rapid and Visual Detection of Trichinella Spp. Using a Lateral Flow Strip Based Recombinase Polymerase Amplification (LF RPA) Assay
abstract: Trichinella spp. are amongst the most widespread parasitic nematodes, primarily live in the muscles of a wide range of vertebrate animals and humans. Human infection occurs by ingestion of raw or undercooked meat containing Trichinella larvae. Accurate diagnosis of Trichinella spp. infection in domestic animals is crucial for the effective prevention and control of human trichinellosis. In the present study, a simple, rapid and accurate diagnostic assay was developed combining recombinase polymerase amplification and a lateral flow strip (LF RPA) to detect Trichinella spp. infection. The LF RPA assay targets Trichinella spp. mitochondrial small subunit ribosomal RNA (rrnS) gene and can detect as low as 100 fg DNA of Trichinella strains, which was approximately 10 times more sensitive than a conventional PCR assay. The LF RPA assay can be performed within 10 25 min, at a wide range of temperatures (25 45degC) and showed no cross reactivity with DNA of other parasites and related host species of Trichinella. The performance of the LF RPA assay in the presence of high concentration of PCR inhibitor was better than that of a conventional PCR assay. Results obtained by LF RPA assay for the detection of experimentally infected mice were comparable to the results obtained by using a conventional PCR, achieving 100% specificity and high sensitivity. These results present the developed LF RPA assay as a new simple, specific, sensitive, rapid and convenient method for the detection of Trichinella infection in domestic animals.

title: Sensing DNA Damage Through ATRIP Recognition of RPA ssDNA Complexes
abstract: The function of the ATR (ataxia telangiectasia mutated and Rad3 related) ATRIP (ATR interacting protein) protein kinase complex is crucial for the cellular response to replication stress and DNA damage. Here, we show that replication protein A (RPA) a protein complex that associates with single stranded DNA (ssDNA) is required for the recruitment of ATR to sites of DNA damage and for ATR mediated Chk1 activation in human cells. In vitro, RPA stimulates the binding of ATRIP to ssDNA. The binding of ATRIP to RPA coated ssDNA enables the ATR ATRIP complex to associate with DNA and stimulates phosphorylation of the Rad17 protein that is bound to DNA. Furthermore, Ddc2, the budding yeast homolog of ATRIP, is specifically recruited to double strand DNA breaks in an RPA dependent manner. A checkpoint deficient mutant of RPA, rfa1 t11, is defective for recruiting Ddc2 to ssDNA both in vivo and in vitro. Our data suggest that RPA coated ssDNA is the critical structure at sites of DNA damage that recruits the ATR ATRIP complex and facilitates its recognition of substrates for phosphorylation and the initiation of checkpoint signaling.

title: EXOSC10 is required for RPA assembly and controlled DNA end resection at DNA double strand breaks
abstract: The exosome is a ribonucleolytic complex that plays important roles in RNA metabolism. Here we show that the exosome is necessary for the repair of DNA double strand breaks (DSBs) in human cells and that RNA clearance is an essential step in homologous recombination. Transcription of DSB flanking sequences results in the production of damage induced long non coding RNAs (dilncRNAs) that engage in DNA RNA hybrid formation. Depletion of EXOSC10, an exosome catalytic subunit, leads to increased dilncRNA and DNA RNA hybrid levels. Moreover, the targeting of the ssDNA binding protein RPA to sites of DNA damage is impaired whereas DNA end resection is hyper stimulated in EXOSC10 depleted cells. The DNA end resection deregulation is abolished by transcription inhibitors, and RNase H1 overexpression restores the RPA recruitment defect caused by EXOSC10 depletion, which suggests that RNA clearance of newly synthesized dilncRNAs is required for RPA recruitment, controlled DNA end resection and assembly of the homologous recombination machinery.The exosome is a ribonucleolytic complex that plays part in RNA processing and degradation. Here, the authors reveal an RNA clearance event in homologous recombination and show the function of EXOSC10, a component of the exosome, in the homeostasis of RNA molecules synthesized at DSB sites.

title: Dynamics and Selective Remodeling of the DNA Binding Domains of RPA
abstract: Replication protein A (RPA) coordinates important DNA metabolic events by stabilizing single stranded DNA (ssDNA) intermediates, activating the DNA damage response and handing off ssDNA to the appropriate downstream players. Six DNA binding domains (DBDs) in RPA promote high affinity binding to ssDNA yet also allow RPA displacement by lower affinity proteins. We generated fluorescent versions of Saccharomyces cerevisiae RPA and visualized the conformational dynamics of individual DBDs in the context of the full length protein. We show that both DBD A and DBD D rapidly bind to and dissociate from ssDNA while RPA remains bound to ssDNA. The recombination mediator protein Rad52 selectively modulates the dynamics of DBD D. These findings reveal how RPA interacting proteins with lower ssDNA binding affinities can access the occluded ssDNA and remodel individual DBDs to replace RPA.A combination of bulk and single molecule fluorescence analysis reveals the choreography of binding and rearrangement of individual DNA binding domains of RPA during homologous recombination.

title: Dual functions for the ssDNA binding protein RPA in meiotic recombination
abstract: Meiotic recombination permits exchange of genetic material between homologous chromosomes. The replication protein A (RPA) complex, the predominant ssDNA binding complex, is required for nearly all aspects of DNA metabolism, but its role in mammalian meiotic recombination remains unknown due to the embryonic lethality of RPA mutant mice. RPA is a heterotrimer of RPA1, RPA2, and RPA3. We find that loss of RPA1, the largest subunit, leads to disappearance of RPA2 and RPA3, resulting in the absence of the RPA complex. Using an inducible germline specific inactivation strategy, we find that loss of RPA completely abrogates loading of RAD51/DMC1 recombinases to programmed meiotic DNA double strand breaks, thus blocking strand invasion required for chromosome pairing and synapsis. Surprisingly, loading of MEIOB, SPATA22, and ATR to DNA double strand breaks is RPA independent and does not promote RAD51/DMC1 recruitment in the absence of RPA. Finally, inactivation of RPA reduces crossover formation. Our results demonstrate that RPA plays two distinct roles in meiotic recombination: an essential role in recombinase recruitment at early stages and an important role in promoting crossover formation at later stages.

title: RPA Phosphorylation Inhibits DNA Resection.
abstract: Genetic recombination in all kingdoms of life initiates when helicases and nucleases process (resect) the free DNA ends to expose single stranded DNA (ssDNA) overhangs. Resection regulation in bacteria is programmed by a DNA sequence, but a general mechanism limiting resection in eukaryotes has remained elusive. Using single molecule imaging of reconstituted human DNA repair factors, we identify phosphorylated RPA (pRPA) as a negative resection regulator. Bloom's syndrome (BLM) helicase together with exonuclease 1 (EXO1) and DNA2 nucleases catalyze kilobase length DNA resection on nucleosome coated DNA. The resulting ssDNA is rapidly bound by RPA, which further stimulates DNA resection. RPA is phosphorylated during resection as part of the DNA damage response (DDR) Remarkably, pRPA inhibits DNA resection in cellular assays and in vitro via inhibition of BLM helicase. pRPA suppresses BLM initiation at DNA ends and promotes the intrinsic helicase strand switching activity. These findings establish that pRPA provides a feedback loop between DNA resection and the DDR.

title: RPA and RAD51: fork reversal, fork protection, and genome stability
abstract: Replication protein A (RPA) and RAD51 are DNA binding proteins that help maintain genome stability during DNA replication. These proteins regulate nucleases, helicases, DNA translocases, and signaling proteins to control replication, repair, recombination, and the DNA damage response. Their different DNA binding mechanisms, enzymatic activities, and binding partners provide unique functionalities that cooperate to ensure that the appropriate activities are deployed at the right time to overcome replication challenges. Here we review and discuss the latest discoveries of the mechanisms by which these proteins work to preserve genome stability, with a focus on their actions in fork reversal and fork protection.Bhat and Cortez discuss current knowledge on the multiple mechanisms by which RPA and RAD51 contribute to genome stability during DNA replication, in particular for replication fork reversal and fork protection.

title: Applying robotic process automation (RPA) in auditing: A framework
abstract: Robotic process automation (RPA) has been widely adopted in many industries, including the accounting industry, to automate well defined and repetitive tasks; however, its application to auditing has lagged behind because of the unique nature of this industry. This study applies RPA in the auditing area. An RPA framework is proposed that frees auditors from doing repetitive and low judgment audit tasks and enables them to focus on tasks that require professional judgment. This paper also demonstrates the feasibility of RPA by implementing a pilot project that applies RPA to the confirmation process.

title: Replication protein A, the laxative that keeps DNA regular: The importance of RPA phosphorylation in maintaining genome stability.
abstract: The eukaryotic ssDNA binding protein, Replication protein A (RPA) was first discovered almost three decades ago. Since then, much progress has been made to elucidate the critical roles for RPA in DNA metabolic pathways that help promote genomic stability. The canonical RPA heterotrimer (RPA1 3) is an essential coordinator of DNA metabolism that interacts with ssDNA and numerous protein partners to coordinate its roles in DNA replication, repair, recombination and telomere maintenance. An alternative form of RPA, termed aRPA, is formed by a complex of RPA4 with RPA1 and RPA3. aRPA is expressed differentially in cells compared to canonical RPA and has been shown to inhibit canonical RPA function while allowing for regular maintenance of cell viability. Interestingly, while aRPA is defective in DNA replication and cell cycle progression, it was shown to play a supporting role in nucleotide excision repair and recombination. The binding domains of canonical RPA interact with a growing number of partners involved in numerous genome maintenance processes. The protein interactions of the RPA ssDNA complex are not only governed by competition between the binding proteins but also by post translation modifications such as phosphorylation. Phosphorylation of RPA2 is an important post translational modification of the RPA complex, and is essential for directing context specific functions of the RPA complex in the DNA damage response. Due to the importance of RPA in cellular metabolism, it was identified as an appealing target for chemotherapeutic drug development that could be used in future cancer treatment regimens.