HuR interacts with human immunodeficiency virus type 1 reverse transcriptase, and modulates reverse transcription in infected cells

Reverse transcription of the genetic material of human immunodeficiency virus type 1 (HIV-1) is a critical step in the replication cycle of this virus. This process, catalyzed by reverse transcriptase (RT), is well characterized at the biochemical level. However, in infected cells, reverse transcription occurs in a multiprotein complex – the reverse transcription complex (RTC) – consisting of viral genomic RNA associated with viral proteins (including RT) and, presumably, as yet uncharacterized cellular proteins. Very little is known about the cellular proteins interacting with the RTC, and with reverse transcriptase in particular. We report here that HIV-1 reverse transcription is affected by the levels of a nucleocytoplasmic shuttling protein – the RNA-binding protein HuR. A direct protein-protein interaction between RT and HuR was observed in a yeast two-hybrid screen and confirmed in vitro by homogenous time-resolved fluorescence (HTRF). We mapped the domain interacting with HuR to the RNAse H domain of RT, and the binding domain for RT to the C-terminus of HuR, partially overlapping the third RRM RNA-binding domain of HuR. HuR silencing with specific siRNAs greatly impaired early and late steps of reverse transcription, significantly inhibiting HIV-1 infection. Moreover, by mutagenesis and immunoprecipitation studies, we could not detect the binding of HuR to the viral RNA. These results suggest that HuR may be involved in and may modulate the reverse transcription reaction of HIV-1, by an as yet unknown mechanism involving a protein-protein interaction with HIV-1 RT

Anisotropy Dependence of Irreversible Switching in Fe/SmCo and FeNi/FePt Exchange Spring Magnet Films

Magnetization reversal in exchange-spring magnet films has been investigated by a First Order Reversal Curve (FORC) technique and vector magnetometry. In Fe/epitaxial-SmCo films, the reversal proceeds by a reversible rotation of the Fe soft layer, followed by an irreversible switching of the SmCo hard layer. The switching fields are clearly manifested by separate steps in both longitudinal and transverse hysteresis loops, as well as sharp boundaries in the FORC distribution. In FeNi/polycrystalline-FePt films, particularly with thin FeNi, the switching fields are masked by the smooth and step-free major loop. However, the FORC diagram still displays a distinct onset of irreversible switching and transverse hysteresis loops exhibit a pair of peaks, whose amplitude is larger than the maximum possible contribution from the FeNi layer alone. This suggests that the FeNi and FePt layers reverse in a continuous process via a vertical spiral. The successive vs. continuous rotation of the soft/hard layer system is primarily due to the different crystal structure of the hard layer, which results in different anisotropies.Comment: 13 pages, 2 figures, APL in pres

A statistical analysis of time trends in atmospheric ethane

Ethane is the most abundant non-methane hydrocarbon in the Earth's atmosphere and an important precursor of tropospheric ozone through various chemical pathways. Ethane is also an indirect greenhouse gas (global warming potential), influencing the atmospheric lifetime of methane through the consumption of the hydroxyl radical (OH). Understanding the development of trends and identifying trend reversals in atmospheric ethane is therefore crucial. Our dataset consists of four series of daily ethane columns obtained from ground-based FTIR measurements. As many other decadal time series, our data are characterized by autocorrelation, heteroskedasticity, and seasonal effects. Additionally, missing observations due to instrument failure or unfavorable measurement conditions are common in such series. The goal of this paper is therefore to analyze trends in atmospheric ethane with statistical tools that correctly address these data features. We present selected methods designed for the analysis of time trends and trend reversals. We consider bootstrap inference on broken linear trends and smoothly varying nonlinear trends. In particular, for the broken trend model, we propose a bootstrap method for inference on the break location and the corresponding changes in slope. For the smooth trend model we construct simultaneous confidence bands around the nonparametrically estimated trend. Our autoregressive wild bootstrap approach, combined with a seasonal filter, is able to handle all issues mentioned above

Spin-density-wave antiferromagnetism of Cr in Fe/Cr(001) superlattices

The antiferromagnetic spin-density-wave (SDW) order of Cr layers in Fe/Cr(001) superlattices was investigated by neutron scattering. For Cr thickness 51-190 {Angstrom}, a transverse SDW is formed for all temperatures below Neel temperature with a single wavevector Q normal to the layers. A coherent magnetic structure forms with the nodes of the SDW near the Fe-Cr interfaces. For thinner Cr layers, the magnetic scattering can be described by commensurate antiferromagnetic order

Cell-Free and Yeast-Based Production of the Malarial Lactate Transporter, PfFNT, Delivers Comparable Yield and Protein Quality

Cell-free protein production is an attractive alternative to cell-based expression. Rapid results, small-volume reactions, irrelevance of protein toxicity, flexibility, and openness of the system are strong points in favor of the cell-free system. However, the in vitro situation lacks the cellular quality control machinery comprising e.g., the translocon for inserting membrane proteins into lipid bilayers, and chaperon-assisted protein degradation pathways. Here, we compare yield and protein quality of the lactate transporter, PfFNT, from malaria parasites when produced in Pichia pastoris yeast, or in an Escherichia coli S30-extract-based cell-free system. Besides solubilization and correct folding, PfFNT requires oligomerization into homopentamers. We assessed PfFNT folding/oligomerization and function by transmission electron microscopy imaging, transport assays, and binding of small-molecule inhibitors. For the latter, we used chromatography of the PfFNT-inhibitor complex with dual-wavelength detection, and biolayer interferometry. Our data show, that PfFNT possesses an intrinsic capability for assuming the correct fold, oligomerization pattern, and functionality during in vitro translation. This competence depended on the detergent present in the cell-free reaction. The choice of detergent further affected purification and inhibitor binding. In conclusion, in the presence of a suitable detergent, cell-free systems are very well capable of producing high quality membrane proteins

Magnetic phase transitions in epitaxial Fe/Cr superlattices

The surface spin-flop and Neel transitions are examined in Fe/Cr superlattices. The surface spin-flop, originally predicted by Mills [Phys. Rev. Lett. 20, 18 (1968)], is observed in Fe/Cr(211) superlattices with antiferromagnetic interlayer coupling and uniaxial in-plane-anisotropy. The Neel transition (T{sub N}) of Cr is observed in Fe/Cr(001) superlattices, for which the onset of antiferromagnetism is at a thickness t{sub Cr} of 42{angstrom}. The bulk value of T{sub N} is approached asymptotically as t{sub Cr} increases and is characterized by a three-dimensional shift exponent. These T{sub N} results are attributed to finite-size effects and spin-frustration near rough Fe-Cr interfaces

Magnetic Phase Transitions in Epitaxial Fe/Cr Superlattices

The surface spin-flop and Neel transitions are examined in Fe/Cr superlattices. The surface spin-flop, originally predicted by Mills [Phys. Rev. Lett. 20, 18 (1968)], is observed in Fe/Cr(211) superlattices with antiferromagnetic interlayer coupling and uniaxial in-plane-anisotropy. The Neel transition (T{sub N}) of Cr is observed in Fe/Cr(001) superlattices, for which the onset of antiferromagnetism is at a thickness t{sub Cr} of 42{angstrom}. The bulk value of T{sub N} is approached asymptotically as t{sub Cr} increases and is characterized by a three-dimensional shift exponent. These T{sub N} results are attributed to finite-size effects and spin-frustration near rough Fe-Cr interfaces