The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.This research was supported by USPHS grant NS073976 (TKH) and P30 ES 06676 that support the NIEHS Center Cell Biology Core and Molecular Genomics Core of UTMB’s NIEHS Center for DNA sequencing. TKP is supported by CA129537 and CA154320. This work was also supported by Fundação para a Ciência e Tecnologia through the project [PTDC/SAU-GMG/101572/2008] and through fellowships [SFRH/BPD/91562/2012 to ASF, SFRH/BD/51059/2010 to ANC]. IB is supported by NIEHS R01 ES018948 and NIAID/AI06288

Circulating microparticles: square the circle

Background: The present review summarizes current knowledge about microparticles (MPs) and provides a systematic overview of last 20 years of research on circulating MPs, with particular focus on their clinical relevance. Results: MPs are a heterogeneous population of cell-derived vesicles, with sizes ranging between 50 and 1000 nm. MPs are capable of transferring peptides, proteins, lipid components, microRNA, mRNA, and DNA from one cell to another without direct cell-to-cell contact. Growing evidence suggests that MPs present in peripheral blood and body fluids contribute to the development and progression of cancer, and are of pathophysiological relevance for autoimmune, inflammatory, infectious, cardiovascular, hematological, and other diseases. MPs have large diagnostic potential as biomarkers; however, due to current technological limitations in purification of MPs and an absence of standardized methods of MP detection, challenges remain in validating the potential of MPs as a non-invasive and early diagnostic platform. Conclusions: Improvements in the effective deciphering of MP molecular signatures will be critical not only for diagnostics, but also for the evaluation of treatment regimens and predicting disease outcomes

Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles

Release of membrane vesicles, a process conserved in both prokaryotes and eukaryotes, represents an evolutionary link, and suggests essential functions of a dynamic extracellular vesicular compartment (including exosomes, microparticles or microvesicles and apoptotic bodies). Compelling evidence supports the significance of this compartment in a broad range of physiological and pathological processes. However, classification of membrane vesicles, protocols of their isolation and detection, molecular details of vesicular release, clearance and biological functions are still under intense investigation. Here, we give a comprehensive overview of extracellular vesicles. After discussing the technical pitfalls and potential artifacts of the rapidly emerging field, we compare results from meta-analyses of published proteomic studies on membrane vesicles. We also summarize clinical implications of membrane vesicles. Lessons from this compartment challenge current paradigms concerning the mechanisms of intercellular communication and immune regulation. Furthermore, its clinical implementation may open new perspectives in translational medicine both in diagnostics and therapy

Precise measurement of the Ds+D^+_s lifetime at Belle II

We measure the lifetime of the $D_s^+$ meson using a data sample of 207 fb$^{-1}$ collected by the Belle II experiment running at the SuperKEKB asymmetric-energy $e^+ e^-$ collider. The lifetime is determined by fitting the decay-time distribution of a sample of $116\times 10^3$ $D_s^+\rightarrow\phi\pi^+$ decays. Our result is \tau^{}_{D^+_s} = (498.7\pm 1.7\,^{+1.1}_{-0.8}) fs, where the first uncertainty is statistical and the second is systematic. This result is significantly more precise than previous measurements.Comment: 7 pages, 4 figures, to be submitted to Physical Review Letter

Search for an invisible Z′Z^\prime in a final state with two muons and missing energy at Belle II

The $L_{\mu}-L_{\tau}$ extension of the standard model predicts the existence of a lepton-flavor-universality-violating $Z^{\prime}$ boson that couples only to the heavier lepton families. We search for such a $Z^\prime$ through its invisible decay in the process $e^+ e^- \to \mu^+ \mu^- Z^{\prime}$. We use a sample of electron-positron collisions at a center-of-mass energy of 10.58GeV collected by the Belle II experiment in 2019-2020, corresponding to an integrated luminosity of 79.7fb$^{-1}$. We find no excess over the expected standard-model background. We set 90$\%$-confidence-level upper limits on the cross section for this process as well as on the coupling of the model, which ranges from $3 \times 10^{-3}$ at low $Z^{\prime}$ masses to 1 at $Z^{\prime}$ masses of 8$GeV/c^{2}$

Precise Measurement of the D0^{0} and D+^{+} Lifetimes at Belle II

We report a measurement of the D$^{0}$ and D$^{+}$ lifetimes using D$^{0}$→K$^{-}$π$^{+}$ and D$^{+}$→K$^{-}$π$^{+}$π$^{+}$ decays reconstructed in e$^{+}$e$^{-}$→$\overline{cc}$ data recorded by the Belle II experiment at the SuperKEKB asymmetric-energy e$^{+}$e$^{-}$ collider. The data, collected at center-of-mass energies at or near the Υ(4S) resonance, correspond to an integrated luminosity of 72 fb$^{-1}$. The results, τ(D$^{0}$)=410.5±1.1(stat)±0.8(syst)  fs and τ(D$^{+}$)=1030.4±4.7(stat)±3.1(syst) fs, are the most precise to date and are consistent with previous determinations

Measurement of branching fractions and direct CPCP asymmetries for B→KπB \to K\pi and B→ππB\to\pi\pi decays at Belle II

We report measurements of the branching fractions and direct $\it{CP}$ asymmetries of the decays $B^0 \to K^+ \pi^-$, $B^+ \to K^+ \pi^0$, $B^+ \to K^0 \pi^+$, and $B^0 \to K^0 \pi^0$, and use these for testing the standard model through an isospin-based sum rule. In addition, we measure the branching fraction and direct $\it{CP}$ asymmetry of the decay $B^+ \to \pi^+\pi^0$ and the branching fraction of the decay $B^0 \to \pi^+\pi^-$. The data are collected with the Belle II detector from $e^+e^-$ collisions at the $\Upsilon(4S)$ resonance produced by the SuperKEKB asymmetric-energy collider and contain $387\times 10^6$ bottom-antibottom meson pairs. Signal yields are determined in two-dimensional fits to background-discriminating variables, and range from 500 to 3900 decays, depending on the channel. We obtain $-0.03 \pm 0.13 \pm 0.04$ for the sum rule, in agreement with the standard model expectation of zero and with a precision comparable to the best existing determinations

Measurement of C ⁣PC\!P asymmetries and branching-fraction ratios for B±→DK±B^\pm \to DK^\pm and Dπ±D\pi^\pm with D→KS0K±π∓D\to K^0_{\rm S} K^\pm\pi^\mp using Belle and Belle II data

We measure $C\!P$ asymmetries and branching-fraction ratios for $B^\pm \to DK^\pm$ and $D\pi^\pm$ decays with $D\to K^0_{\rm S} K^\pm\pi^\mp$, where $D$ is a superposition of $D^0$ and $\bar{D}^0$. We use the full data set of the Belle experiment, containing $772\times 10^6~B\bar{B}$ pairs, and data from the Belle~II experiment, containing $387\times 10^6~B\bar{B}$ pairs, both collected in electron-positron collisions at the $\Upsilon(4S)$ resonance. Our results provide model-independent information on the unitarity triangle angle $\phi_3$.Comment: 26 pages, 8 figure