Novel inhibition mechanism and potent antiviral activity of translocation-deficient reverse transcriptase inhibitors [abstract]

Abstract only availableNucleoside RT inhibitors (NRTIs) are among the most potent anti-HIV agents and act as chain terminators because they lack a 3'OH. However, this feature can reduce affinity for RT compared to the analogous dNTP substrate, as well as reduced intracellular conversion to the active dNTP. To overcome this, it was shown that certain nucleosides that retain the 3'OH and have substitutions at the 4' ribose and 2 position of the base have exceptional antiviral properties. One of these compounds, 4'-ethynyl, 2-fluoro deoxy-adenosine (4'E-2FdA) is the most potent NRTI inhibitor against wild-type and multi-drug resistant HIV viruses described to date. We have recently reported that 4'E-2FdA acts as a chain terminator despite the presence of an accessible 3'OH. We show that after 4'E-2FdA-MP incorporation, RT does not bind the next incoming dNTP. We analyzed RT translocation on different sequences terminated with 4'E-2FdA-MP, and found that even at sequences when RT is naturally found post-translocated, the inhibitor prevents translocation. This decrease in translocation efficiency explains the reduced binding of the next incoming dNTP and the termination of elongation. While the inhibitor stabilizes the pre-translocated 4'E-2FdA-MP-terminated primer, the pyrophosphate-dependent excision rate of 4'E-2FdA-MP was not very high compared to ddAMP. In conclusion, this highly potent chain termination activity arises from difficulty of the primer 3'-terminus to translocate following incorporation of the compound, and not from simple steric hindrance due to the 4' substitution. Therefore, we propose that 4'E-2FdA is a Translocation-Deficient Reverse Transcriptase Inhibitor (TDRTI) that acts by a novel mechanism.NIH grant to S. Sarafiano

Analytic Solutions of The Wheeler-DeWitt Equation in Spherically Symmetric Space-time

We study the quantum theory of the Einstein-Maxwell action with a cosmological term in the spherically symmetric space-time, and explored quantum black hole solutions in Reissner-Nordstrom-de Sitter geometry. We succeeded to obtain analytic solutions to satisfy both the energy and momentum constraints.Comment: LaTeX file, 15 page

Bioorganic synthesis of a recombinant HIV-1 fusion inhibitor, SC35EK, with an N-terminal pyroglutamate capping group.

The bioorganic synthesis of an end-capped anti-HIV peptide from a recombinant protein was investigated. Cyanogen bromide-mediated cleavage of two Met-Gln sites across the target anti-HIV sequence generated an HIV-1 fusion inhibitor (SC35EK) analog bearing an N-terminal pyroglutamate (pGlu) residue and a C-terminal homoserine lactone (Hsl) residue. The end-capped peptide, pGlu-SC35EK-Hsl, had similar bioactivity and biophysical properties to the parent peptide, and an improved resistance to peptidase-mediated degradation was observed compared with the non-end-capped peptide obtained using standard recombinant technology

Discovery of a Giant Lyα Emitter Near the Reionization Epoch

‘In these times, during the rise in the popularity of institutional repositories, the Society does not forbid authors from depositing their work in such repositories. However, the AAS regards the deposit of scholarly work in such repositories to be a decision of the individual scholar, as long as the individual's actions respect the diligence of the journals and their reviewers.’ Original article can be found at : http://iopscience.iop.org/ Copyright American Astronomical SocietyWe report the discovery of a giant Lyα emitter (LAE) with a Spitzer/Infrared Array Camera (IRAC) counterpart near the reionization epoch at z = 6.595. The giant LAE is found from the extensive 1 deg2 Subaru narrowband survey for z = 6.6 LAEs in the Subaru/XMM-Newton Deep Survey (SXDS) field, and subsequently identified by deep spectroscopy of Keck/DEIMOS and Magellan/IMACS. Among our 207 LAE candidates, this LAE is not only the brightest narrowband object with L(Lyα) = 3.9 ± 0.2 × 1043 erg s–1 in our survey volume of 106 Mpc3, but also a spatially extended Lyα nebula with the largest isophotal area whose major axis is at least 3''. This object is more likely to be a large Lyα nebula with a size of 17 kpc than to be a strongly lensed galaxy by a foreground object. Our Keck spectrum with medium-high spectral and spatial resolutions suggests that the velocity width is v FWHM = 251 ± 21 km s–1, and that the line-center velocity changes by 60 km s–1 in a 10 kpc range. The stellar mass and star formation rate are estimated to be 0.9-5.0 × 1010 M and >34 M yr–1, respectively, from the combination of deep optical to infrared images of Subaru, UKIDSS-Ultra Deep Survey, and Spitzer/IRAC. Although the nature of this object is not yet clearly understood, this could be an important object for studying cooling clouds accreting onto a massive halo, or forming-massive galaxies with significant outflows contributing to cosmic reionization and metal enrichment of intergalactic medium.Peer reviewe

Identification of minimal sequence for HIV-1 fusion inhibitors.

Emergence of multi-drug resistant HIV-1 is a serious problem for AIDS treatment. Recently, the virus-cell membrane fusion process has been identified as a promising target for the development of novel drugs against these resistant variants. In this study, we identified a 29-residue peptide fusion inhibitor, SC29EK, which shows activity comparable to the previously reported inhibitor SC35EK. Some residues in SC29EK not required for interaction with virus gp41 heptad repeat 1 (HR1) were replaced with a non-proteinogenic amino acid, 2-aminoisobutyric acid (Aib), to stabilize the alpha-helix structure and to provide resistance to peptidases

ALMA twenty-six arcmin2^2 survey of GOODS-S at one-millimeter (ASAGAO): Near-infrared-dark faint ALMA sources

We report detections of two 1.2 mm continuum sources ($S_\mathrm{1.2mm}$ ~ 0.6 mJy) without any counterparts in the deep $H$- and/or $K$-band image (i.e., $K$-band magnitude $\gtrsim$ 26 mag). These near-infrared-dark faint millimeter sources are uncovered by ASAGAO, a deep and wide-field ($\simeq$ 26 arcmin$^2$) Atacama Large Millimeter/submillimeter Array (ALMA) 1.2 mm survey. One has a red IRAC (3.6 and 4.5 $\mu$m) counterpart, and the other has been independently detected at 850 and 870 $\mu$m using SCUBA2 and ALMA Band 7, respectively. Their optical to radio spectral energy distributions indicate that they can lie at $z \gtrsim$ 3-5 and can be in the early phase of massive galaxy formation. Their contribution to the cosmic star formation rate density is estimated to be ~ 1 $\times$ 10$^{-3}$ $M_\odot$ yr$^{-1}$ Mpc$^{-3}$ if they lie somewhere in the redshift range of $z$ ~ 3-5. This value can be consistent with, or greater than that of bright submillimeter galaxies ($S_\mathrm{870\mu m}>$ 4.2 mJy) at $z$ ~ 3-5. We also uncover 3 more candidates near-infrared-dark faint ALMA sources without any counterparts ($S_\mathrm{1.2mm}$ ~ 0.45-0.86 mJy). These results show that an unbiased ALMA survey can reveal the dust-obscured star formation activities, which were missed in previous deep optical/near-infrared surveys.Comment: 10 pages, 6 figures, accepted for publication in Ap

K70Q Adds High-Level Tenofovir Resistance to “Q151M Complex” HIV Reverse Transcriptase through the Enhanced Discrimination Mechanism

HIV-1 carrying the “Q151M complex” reverse transcriptase (RT) mutations (A62V/V75I/F77L/F116Y/Q151M, or Q151Mc) is resistant to many FDA-approved nucleoside RT inhibitors (NRTIs), but has been considered susceptible to tenofovir disoproxil fumarate (TFV-DF or TDF). We have isolated from a TFV-DF-treated HIV patient a Q151Mc-containing clinical isolate with high phenotypic resistance to TFV-DF. Analysis of the genotypic and phenotypic testing over the course of this patient's therapy lead us to hypothesize that TFV-DF resistance emerged upon appearance of the previously unreported K70Q mutation in the Q151Mc background. Virological analysis showed that HIV with only K70Q was not significantly resistant to TFV-DF. However, addition of K70Q to the Q151Mc background significantly enhanced resistance to several approved NRTIs, and also resulted in high-level (10-fold) resistance to TFV-DF. Biochemical experiments established that the increased resistance to tenofovir is not the result of enhanced excision, as K70Q/Q151Mc RT exhibited diminished, rather than enhanced ATP-based primer unblocking activity. Pre-steady state kinetic analysis of the recombinant enzymes demonstrated that addition of the K70Q mutation selectively decreases the binding of tenofovir-diphosphate (TFV-DP), resulting in reduced incorporation of TFV into the nascent DNA chain. Molecular dynamics simulations suggest that changes in the hydrogen bonding pattern in the polymerase active site of K70Q/Q151Mc RT may contribute to the observed changes in binding and incorporation of TFV-DP. The novel pattern of TFV-resistance may help adjust therapeutic strategies for NRTI-experienced patients with multi-drug resistant (MDR) mutations