Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of disease 2019 (COVID-19). SARS-CoV-2 a single-stranded positive-sense RNA virus. Like other coronaviruses, has an unusually large genome that encodes four structural proteins and sixteen nonstructural proteins. The nucleocapsid phosphoprotein N essential for linking viral to membrane. Both N-terminal binding (N-NTD) C-terminal dimerization domains are involved in capturing and, intrinsically disordered...

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of COVID-19 pandemic. 2'-O-RNA methyltransferase (MTase) one enzymes this virus that a potential target for antiviral therapy as it crucial RNA cap formation; an essential process viral stability. This MTase function associated with nsp16 protein, which requires cofactor, nsp10, its proper activity. Here we show crystal structure nsp10-nsp16 complex bound to pan-MTase inhibitor sinefungin in active site. Our structural...

SLX4 binds to three nucleases (XPF-ERCC1, MUS81-EME1, and SLX1), its deficiency leads genomic instability, sensitivity DNA crosslinking agents, Fanconi anemia. However, it is not understood how associated act in crosslink repair. Here, we uncover consequences of mouse Slx4 reveal function Slx4-deficient mice develop epithelial cancers have a contracted hematopoietic stem cell pool. The N-terminal domain (mini-SLX4) that only XPF-ERCC1 sufficient confer resistance agents. Recombinant...

Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of Coronavirus disease 2019 (COVID-19) which currently negatively affecting population and disrupting global economy. SARS-CoV-2 belongs to +RNA virus family that utilize single-stranded positive-sense RNA molecules as genomes. SARS-CoV-2, like other coronaviruses, has an unusually large genome for a encodes four structural proteins – matrix (M), small envelope (E), spike (S) nucleocapsid...

Monkeypox is a disease with pandemic potential. It caused by the monkeypox virus (MPXV), double-stranded DNA from Poxviridae family, that replicates in cytoplasm and must encode for its own RNA processing machinery including capping machinery. Here, we present crystal structures of 2'-O-RNA methyltransferase (MTase) VP39 complex pan-MTase inhibitor sinefungin series inhibitors were discovered based on it. A comparison this MTase enzymes unrelated single-stranded viruses (SARS-CoV-2 Zika)...

Coronaviral methyltransferases (MTases), nsp10/16 and nsp14, catalyze the last two steps of viral RNA-cap creation that takes place in cytoplasm. This cap is essential for stability RNA and, most importantly, evasion innate immune system. Non-capped recognized by immunity which leads to its degradation activation antiviral immunity. As a result, both coronaviral MTases are center scientific scrutiny. Recently, X-ray cryo-EM structures enzymes were solved even complex with other parts...

FoxO4 belongs to the "O" subset of forkhead transcription factors, which participate in various cellular processes. The DNA binding domain (DBD) consists three-helix bundle resting on a small antiparallel β-sheet from two extended loops protrude and create wing-like structures. wing W2 FoxO factors contains 14-3-3 protein-binding motif that is phosphorylated by protein kinase B response insulin or growth factors. In this report, we investigated role N-terminal loop (portion located upstream...

The role of 14-3-3 proteins in the regulation FOXO forkhead transcription factors is at least 2-fold. First, binding inhibits interaction between and target DNA. Second, prevent nuclear reimport by masking their localization signal. exact mechanisms these processes are still unclear, mainly due to lack structural data. In this work, we used fluorescence spectroscopy investigate mechanism protein-dependent inhibition FOXO4 DNA-binding properties. Time-resolved measurements revealed that...

The RNA 2'-O methyltransferase (MTase) VP39 of the monkeypox virus (MpxV) participates in capping within poxviruses. Sub-micromolar inhibitors targeting this enzyme were already reported. However, these 7-deaza analogs S-adenosyl methionine (SAH) had not been tested cellular assays until now. In study, we employed plaque and cytopathic effect-based to evaluate effectiveness compounds. All compounds demonstrated antiviral activity against MpxV, with EC50 values ranging from 0.06 2.7 μM....

14-3-3 proteins are important regulators of numerous cellular signaling circuits. They bind to phosphorylated protein ligands and regulate their functions by a number different mechanisms. The C-terminal part the is known be involved in regulation binding properties. structure this region unknown; however, possible location stretch within ligand groove has been suggested. To fully understand role properties, we investigated physical its changes upon binding. For purpose, have used Förster...

Tyrosine hydroxylase (TH) catalyzes the first step in biosynthesis of catecholamines. Regulation TH enzyme activity is controlled through posttranslational modification its regulatory domain. The domain can be phosphorylated at four serines (8, 19, 31, and 40) by a variety protein kinases. Phosphorylation Ser19 does not itself increase but induces binding to 14-3-3 protein. That leads enhancement with still fully understood mechanism. main goal this work was investigate whether affects...

Mpox is a zoonotic disease caused by the mpox virus (MPXV), which has gained attention due to its rapid and widespread transmission, with reports from more than 100 countries. The belongs Orthopoxvirus genus, also includes variola vaccinia virus. In poxviruses, RNA cap crucial for translation stability of viral mRNAs immune evasion. This study presents crystal structure 2'-O-methyltransfarase VP39 in complex short cap-0 RNA. substrate binds protein without causing any significant changes...

The 20S proteasome, a critical component of the ubiquitin-proteasome system, plays central role in regulating protein degradation eukaryotic cells. Marizomib (MZB), natural γ-lactam-β-lactone compound derived from Salinispora tropica, is potent proteasome covalent inhibitor with demonstrated anticancer properties. Its broad-spectrum inhibition all three subunits and ability to cross blood-brain barrier has made it promising therapeutic candidate for glioblastoma. Here, we present cryo-EM...

Segments of used drinking water pipes galvanised steel (GS), cross-linked polyethylene (PEX), copper (Cu) or new medium-density (PE) were investigated for the formation biofilm and survival E. coli in phase. Pipes filled with incubated at 15 °C 35 under static conditions. Biofilm was followed during 32, 40 56 (58) d. The most dense formed on GS, reaching approximately 4.7×105 CFU/cm2 measured as heterotrophic plate count (HPC), other materials density reached 3×103 PE PEX 5×101 5×102 Cu...

The 20S proteasome, a critical component of the ubiquitin–proteasome system, plays central role in regulating protein degradation eukaryotic cells. Marizomib (MZB), also known as salinosporamide A, is natural γ-lactam-β-lactone compound derived from Salinispora tropica and potent proteasome covalent inhibitor with demonstrated anticancer properties. Its broad-spectrum inhibition all three subunits its ability to cross blood–brain barrier has made it promising therapeutic candidate for...

Although oxidative stress is a key aspect of innate immunity, little known about how host-restricted pathogens successfully repair DNA damage. Base excision responsible for correcting nucleobases damaged by stress, and essential bloodstream infection caused the human pathogen, Neisseria meningitidis. We have characterized meningococcal base enzymes involved in recognition removal nucleobases, incision backbone. demonstrate that bi-functional glycosylase/lyases Nth MutM share several...

Some coronaviruses are dangerous pathogens, while some cause only common colds. The reasons not understood, although the spike proteins probably play an important role.

Base excision repair (BER) is a highly conserved DNA pathway throughout all kingdoms from bacteria to humans. Whereas several enzymes are required complete the multistep process of damaged bases, apurinic-apyrimidic (AP) endonucleases play an essential role in enabling by recognizing intermediary abasic sites cleaving phosphodiester backbone 5′ site. Despite extensive study, there no structure bacterial AP endonuclease bound substrate DNA. Furthermore, structural mechanism for AP-site...

Bacterial MutM is a DNA repair glycosylase removing damage generated from oxidative stress and, therefore, preventing mutations and genomic instability. belongs to the Fpg/Nei family of prokaryotic enzymes sharing structural functional similarities with their eukaryotic counterparts, for example, NEIL1-NEIL3. Here, we present two crystal structures pathogenic Neisseria meningitidis: holoenzyme bound DNA. The free enzyme exists in an open conformation, while upon binding DNA, both undergo...

Spanish flu, polio epidemics, and the ongoing COVID-19 pandemic are most profound examples of severe widespread diseases caused by RNA viruses. The coronavirus acute respiratory syndrome 2 (SARS-CoV-2) demands affordable reliable assays for testing antivirals. To test inhibitors viral proteases, we have developed an inexpensive high-throughput assay based on fluorescent energy transfer (FRET). We assayed array papain-like protease from SARS-CoV-2 validated it tick-borne encephalitis virus to...