Association of Cystic Fibrosis Transmembrane Conductance Regulator with Epithelial Sodium Channel Subunits Carrying Liddle's Syndrome Mutations

The association of the cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC) in the pathophysiology of cystic fibrosis (CF) is controversial. Previously, we demonstrated a close physical association between wild type (WT) CFTR and WT ENaC. We have also shown that the F508del CFTR fails to associate with ENaC unless the mutant protein is rescued pharmacologically or by low temperature. In this study, we present the evidence for a direct physical association between WT CFTR and ENaC subunits carrying Liddle's syndrome mutations. We show that all three ENaC subunits bearing Liddle's syndrome mutations (both point mutations and the complete truncation of the carboxy terminus), could be co-immunoprecipitated with WT CFTR. The biochemical studies were complemented by fluorescence lifetime imaging microscopy (FLIM), a distance-dependent approach that monitors protein-protein interactions between fluorescently labeled molecules. Our measurements revealed significantly increased FRET between CFTR and all tested ENaC combinations as compared to controls (ECFP and EYFP co-transfected cells). Our findings are consistent with the notion that CFTR and ENaC are within reach of each other even in the setting of Liddle's syndrome mutations, suggestive of a direct intermolecular interaction between these two proteins.

Prespacers formed during primed adaptation associate with the Cas1–Cas2 adaptation complex and the Cas3 interference nuclease–helicase

For Type I CRISPR-Cas systems, a mode of CRISPR adaptation named priming has been described. Priming allows specific and highly efficient acquisition of new spacers from DNA recognized (primed) by the Cascade-crRNA (CRISPR RNA) effector complex. Recognition of the priming protospacer by Cascade-crRNA serves as a signal for engaging the Cas3 nuclease–helicase required for both interference and primed adaptation, suggesting the existence of a primed adaptation complex (PAC) containing the Cas1–Cas2 adaptation integrase and Cas3. To detect this complex in vivo, we here performed chromatin immunoprecipitation with Cas3-specific and Cas1-specific antibodies using cells undergoing primed adaptation. We found that prespacers are bound by both Cas1 (presumably, as part of the Cas1–Cas2 integrase) and Cas3, implying direct physical association of the interference and adaptation machineries as part of PAC.

Compressed magnetized shells of atomic gas and the formation of the Corona Australis molecular cloud

We present the identification of the previously unnoticed physical association between the Corona Australis molecular cloud (CrA), traced by interstellar dust emission, and two shell-like structures observed with line emission of atomic hydrogen (HI) at 21 cm. Although the existence of the two shells had already been reported in the literature, the physical link between the HI emission and CrA had never been highlighted until now. We used both Planck and Herschel data to trace dust emission and the Galactic All Sky HI Survey (GASS) to trace HI. The physical association between CrA and the shells is assessed based both on spectroscopic observations of molecular and atomic gas and on dust extinction data with Gaia. The shells are located at a distance between ~140 and ~190 pc, which is comparable to the distance of CrA, which we derived as (150.5 ± 6.3) pc. We also employed dust polarization observations from Planck to trace the magnetic-field structure of the shells. Both of them show patterns of magnetic-field lines following the edge of the shells consistently with the magnetic-field morphology of CrA. We estimated the magnetic-field strength at the intersection of the two shells via the Davis-Chandrasekhar-Fermi (DCF) method. Despite the many caveats that are behind the DCF method, we find a magnetic-field strength of (27 ± 8) μG, which is at least a factor of two larger than the magnetic-field strength computed off of the HI shells. This value is also significantly larger compared to the typical values of a few μG found in the diffuse HI gas from Zeeman splitting. We interpret this as the result of magnetic-field compression caused by the shell expansion. This study supports a scenario of molecular-cloud formation triggered by supersonic compression of cold magnetized HI gas from expanding interstellar bubbles.

O dwóch podejściach do badań dialektów miejskich w Polsce i Czechach

Two tendencies in researching urban dialects in Poland and the Czech RepublicThis article presents a study in urban dialects of Polish and Czech researchers. The author introduces the history and structural features of Polish and Czech urban dialects, shows differences between them in theoretical, structural and typological aspects. The main differences in the structure are caused by the various differentiation of both languages. The author comes to the conclusion that Czech urban dialects consist of many variants of the national language and are positioned high culturally and socially. Polish urban dialects that appeared in a community of low social status reveal differences in lexicon and phonetics. Nowadays, the urban dialects of both languages function as advertising and an element of folklore and are proof of citizens’ physical association with their cities and show their mentality and cultural characteristics. Два подхода в изучении городских диалектов в Польше и ЧехииСтатья посвящена проблематике изучения городских диалектов польскими и чешскими учеными. Рассматривается история и структурная характеристика польских и чешских городских диалектов. В работе представлены различия языка города в Польше и Чехии в теоретическом, структурном и типологическом планах. Основные несоответствия в структурах городских диалектов объяснены разницами в дифференциации национального языка как польского, так и чешского. Изученные исследования показали, что чешские городские диалекты представляют собой смесь разных вариантов национального языка, а их культурносоциальный статус является достаточно высоким. Польские городские диалекты, возникшие среди носителей с низким социальным статусом, отличаются друд от друга, в них имеются различия на лексическом и фонетическом уровях. В настоящее время городские диалекты обоих языков выполняют фольклорно-рекламную функцию и свидетельствуют о связи горожан со своим городом, их менталитете и культурных особенностях.

Single subunit degradation of WIZ, a lenalidomide- and pomalidomide-dependent substrate of E3 ubiquitin ligase CRL4CRBN

Immunomodulators (IMiDs) are an effective class of drugs used to treat blood cancers. IMiDs are believed to work by recruiting protein targets containing a β-hairpin motif for ubiquitination by E3 ubiquitin ligase complexes composed of cereblon (CRBN), Cullin-4a (CUL4a), DNA Damage Binding protein-1 (DDB1), and Ring Box-1 (RBX1). The ubiquitinated protein is subsequently degraded by the proteasome. By characterizing the repertoire of proteins that show an increased physical association with CRBN after IMiD treatment, we identified a novel IMiD substrate, Widely Interspaced Zinc Finger Motifs (WIZ). WIZ contains a C2H2 zinc finger domain, like several other substrates that were previously characterized. We demonstrate that IMiDs stabilize physical association of WIZ with CRBN, deplete WIZ steady state protein levels in a way that is dependent on E3 ligase activity, and enhance the rate of its degradation. Notably, proteins that assemble with WIZ are co-recruited to CRBN by IMiDs but are not degraded, illustrating the potential of targeted protein degradation to eliminate individual subunits of a protein complex. These findings suggest that systematic characterization of the full repertoire of proteins that are targeted for degradation by IMiD compounds will be required to better understand their biological effects.SynopsisProteolysis Targeting Chimeras (PROTACs) can be used to precisely target a subunit of a transcriptional complex for degradation in subpopulations of cells.

Salicylic acid-driven association of LENRV and NIMIN1/NIMIN2 binding domain regions in the C-terminus of tobacco NPR1 transduces SAR signal

SummaryNONEXPRESSOR OF PATHOGENESIS-RELATED (PR) GENES1 (NPR1) is the central regulator of salicylic acid (SA)-induced PR-1 gene expression and systemic acquired resistance (SAR). The mechanism how SA is transduced through NPR1 is discussed controversially. Previously, we showed that Arabidopsis and tobacco (Nt) NPR1 contain two domains in their C-terminal thirds with relevance to SA signaling. SA sensitivity of NPR1 relies on the arginine residue in the LENRV motif, and SA-induced NIM1-INTERACTING (NIMIN, N) proteins bind to a highly conserved sequence termed N1/N2 binding domain (BD).We demonstrate that LENRV and N1/N2BD regions of tobacco NPR1, separated from each other, interact in yeast, in vitro, in plant and in animal cells. Physical association of LENRV and N1/N2BD parts is enhanced considerably by SA and functional analogs, but not by a non-functional analog. Furthermore, physical association requires R431 and is most effective with intact LENRV and N1/N2BD interfaces.Association of separated LENRV and N1/N2BD parts by SA reconstitutes a functional NtNPR1 C-terminus, displaying transcription activity and able to interact with TGA transcription factors at two distinct sites.Tobacco NIMIN proteins can assemble LENRV and N1/N2BD parts into ternary complexes suggesting that NIMINs shape the NPR1 C-terminus to modulate SA signaling.

Semiotics

As the study of signification, semiotics takes as its central task that of describing how one thing can mean another. Alternatively, since this philosophical problem is also a psychological one, its job could be said to be that of describing how one thing can bring something else to mind, how on seeing ’x’ someone can be induced to think about ’y’ even though ’y’ is absent. A person in whose head ’y’ has been brought to mind may be responding to an ’x’ someone else has transmitted with the intention of its signifying ’y’; or, mistakenly, responding to an ’x’ someone has transmitted in the guileless expectation of its signifying some ’z’; or, often, responding to an ’x’ that comes to his notice without anybody’s apparent intention at all. Words, for example, generally signify because someone intends them to, and ideally (though not always) they signify what is intended; whereas clouds signify – a coming storm, a whale – because we so interpret them, not because they shaped themselves to convey some meaning. Obviously the study of signification forms an integral part of the study of thinking, since no object can itself enter the brain, barring fatal mischance, and so it must be represented by some mental (that is, neural) ’x’ that signifies it. Signifiers are equally essential for creatures far lower than humans, as when a chemical signal ’x’ emitted by some bacterium signifies to one of its colleagues some ‘y’ such as ’there’s a dearth of food hereabouts’. There are a number of ways in which an ’x’ can signify some ’y’, but for humans these are chiefly: by physical association; by physical resemblance; and/or by arbitrary convention. When we take some ’x’ as signifying some ‘y’ we are often guessing; our guess is subject to checking by interpretative (re)appraisal.

Crime and Victimization in Miami

This chapter considers the flipside of globalization and economic progress in Miami—urban crime. Crime is not homogeneous; there are several distinct types, each with its own etiology and consequences. Homicide, for example, is related to but emerges from causal configurations different from those leading to property theft or money laundering. The chapter reviews leading theories of urban crime in order to develop a framework to fit empirical findings. It highlights the fact that not all urban crime occurs at street level or involves personal victims. It indicates the physical association between different forms of crime and locations in urban space. It shows that wealthier areas of town are not crime-free. They are sites for different forms of crime—more complex, apparently “victimless,” but at times more insidious than the street varieties.