Search

U1 small nuclear ribonucleoparticle (U1 snRNP) plays a central role during RNA processing. Previous structures of U1 snRNP revealed how the ribonucleoparticle is organized and recognizes the pre-mRNA substrate at the exon–intron junction. As with many other ribonucleoparticles involved in RNA metabolism, U1 snRNP contains extensions made of low complexity sequences. Here, we developed a protocol to reconstitute U1 snRNP in vitro using mostly full-length components in order to perform liquid-state NMR spectroscopy. The accuracy of the reconstitution was validated by probing the shape and structure of the particle by SANS and cryo-EM. Using an NMR spectroscopy-based approach, we probed, for the first time, the U1 snRNP tails at atomic detail and our results confirm their high degree of flexibility. We also monitored the labile interaction between the splicing factor PTBP1 and U1 snRNP and validated the U1 snRNA stem loop 4 as a binding site for the splicing regulator on the ribonucleoparticle. Altogether, we developed a method to probe the intrinsically disordered regions of U1 snRNP and map the interactions controlling splicing regulation. This approach could be used to get insights into the molecular mechanisms of alternative splicing and screen for potential RNA therapeutics., Nucleic Acids Research, 49 (11), ISSN:1362-4962, ISSN:0301-5610

Power amplifiers have been playing a vital role in most wireless communication systems. In order to improve efficiency of wireless systems, advanced transmitter architectures, such as Doherty amplifiers, outphasing amplifiers, supply voltage modulation techniques are widely used. The goal of this work is to develop novel techniques for building load modulation transmitters based on class-E power amplifiers. The first contribution is an analytical model for derivation load network parameters. The proposed model derives the parameters for both the peak and back-off power levels providing high efficiency. The proposed model demonstrates, that class-E PA with shunt capacitance and shunt filter is capable of providing high drain efficiency for back-off output power levels. The second contribution is a design of a wideband class-E power amplifier (PA) with shunt capacitance and shunt filter. The broadband operation has been achieved by application of the double reactance compensation technique. Simulated and experimental results are presented. The performance of the fabricated PA is compared with existing wideband PAs. The third contribution is application of the proposed technique to outphasing PA design. The designed outphasing PA was optimized, fabricated and tested. A possibility to extend the operational bandwidth of the PA is considered. Also the application of the proposed technique to Doherty PA design is demonstrated. The fourth contribution is linearization of outphasing PA. Firstly, an analytical model describing the nonlinearity of nonisolated combiners under amplitude imbalance is presented. Secondly, a novel phase-only predistortion technique for class-E outphasing PAs is proposed. Thirdly, linearization of the fabricated outphasing PA based on memory polynomial model is demonstrated using a 64QAM OFDM modulated signal with 20 MHz bandwidth. Overall, this work provides novel techniques for load modulation transmitter design based on class-E power amplifiers with shunt capacitance and shunt filter.

The glycoprotein uromodulin (UMOD) is the most abundant protein in human urine and forms filamentous homopolymers that encapsulate and aggregate uropathogens, promoting pathogen clearance by urine excretion. Despite its critical role in the innate immune response against urinary tract infections, the structural basis and mechanism of UMOD polymerization remained unknown. Here, we present the cryo-EM structure of the UMOD filament core at 3.5 Å resolution, comprised of the bipartite zona pellucida (ZP) module in a helical arrangement with a rise of ~65 Å and a twist of ~180°. The immunoglobulin-like ZPN and ZPC subdomains of each monomer are separated by a long linker that interacts with the preceding ZPC and following ZPN subdomains by β-sheet complementation. The unique filament architecture suggests an assembly mechanism in which subunit incorporation could be synchronized with proteolytic cleavage of the C-terminal pro-peptide that anchors assembly-incompetent UMOD precursors to the membrane. ISSN:2050-084X

Therapeutics with novel modes of action and a low risk of generating resistance are urgently needed to combat drug-resistant Plasmodium falciparum malaria. Here, we report that the peptide vinyl sulfones WLL-vs (WLL) and WLW-vs (WLW), highly selective covalent inhibitors of the P. falciparum proteasome, potently eliminate genetically diverse parasites, including K13-mutant, artemisinin-resistant lines, and are particularly active against ring-stage parasites. Selection studies reveal that parasites do not readily acquire resistance to WLL or WLW and that mutations in the β2, β5 or β6 subunits of the 20S proteasome core particle or in components of the 19S proteasome regulatory particle yield only hundred-fold decreases in susceptibility. We observed no cross-resistance between WLL and WLW. Moreover, most mutations that conferred a modest loss of parasite susceptibility to one inhibitor significantly increased sensitivity to the other. These inhibitors potently synergized multiple chemically diverse classes of antimalarial agents, implicating a shared disruption of proteostasis in their modes of action. These results underscore the potential of targeting the Plasmodium proteasome with covalent small molecule inhibitors as a means of combating multidrug-resistant malaria.

Efficient and linear power amplifiers (PA) are an essential part of forthcoming 5G wireless systems. Outphasing class-E PAs offer high power efficiency and an option for higher efficiency cellular networks. However, they employ signal component separators, which split the signal into two paths. In order to efficiently recombine the signal, nonlinear power combiners are used. This paper proposes a novel phase-only predistortion technique for outphasing class-E PAs. The predistortion coefficients can be extracted based on AMAM characteristics of the output signal and an analytical model of an outphasing Class E PA. The suggested technique has been validated by simulation of an outphasing power amplifier in ADS Ptolemy software. It is shown that applying this technique to a 16QAM OFDM modulated signal with 20 MHz bandwidth improves error vector magnitude (EVM) from 10.39% to 2.43% compared to the signal without predistortion.