Noncoding RNAs (ncRNAs) play increasingly appreciated gene-regulatory roles. Here, we describe a regulatory network centered on four ncRNAs—a long ncRNA, a circular RNA, and two microRNAs—using gene ...editing in mice to probe the molecular consequences of disrupting key components of this network. The long ncRNA Cyrano uses an extensively paired site to miR-7 to trigger destruction of this microRNA. Cyrano-directed miR-7 degradation is much more effective than previously described examples of target-directed microRNA degradation, which come primarily from studies of artificial and viral RNAs. By reducing miR-7 levels, Cyrano prevents repression of miR-7–targeted mRNAs and enables accumulation of Cdr1as, a circular RNA known to regulate neuronal activity. Without Cyrano, excess miR-7 causes cytoplasmic destruction of Cdr1as in neurons, in part through enhanced slicing of Cdr1as by a second miRNA, miR-671. Thus, several types of ncRNAs can collaborate to establish a sophisticated regulatory network.
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•A long noncoding RNA, a circular RNA, and two microRNAs form a regulatory network•The Cyrano long noncoding RNA directs potent, multiple-turnover destruction of miR-7•Unchecked miR-7 prevents accumulation of Cdr1as circular RNA in cytoplasm of neurons•miR-7 prevents this accumulation by enhancing the miR-671–directed slicing of Cdr1as
Three different classes of noncoding RNA converge in a regulatory network whereby a long noncoding RNA represses a microRNA via target-directed microRNA degradation, which in turn enables the accumulation of a circular RNA in the mouse brain.
Recent studies have reported that competitive endogenous RNAs (ceRNAs) can act as sponges for a microRNA (miRNA) through their binding sites and that changes in ceRNA abundances from individual genes ...can modulate the activity of miRNAs. Consideration of this hypothesis would benefit from knowing the quantitative relationship between a miRNA and its endogenous target sites. Here, we altered intracellular target site abundance through expression of an miR-122 target in hepatocytes and livers and analyzed the effects on miR-122 target genes. Target repression was released in a threshold-like manner at high target site abundance (≥1.5 × 105 added target sites per cell), and this threshold was insensitive to the effective levels of the miRNA. Furthermore, in response to extreme metabolic liver disease models, global target site abundance of hepatocytes did not change sufficiently to affect miRNA-mediated repression. Thus, modulation of miRNA target abundance is unlikely to cause significant effects on gene expression and metabolism through a ceRNA effect.
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•Target repression is released only after adding many competing target sites•The number of added sites required for derepression is independent of miRNA levels•The apparent abundance of intracellular target sites exceeds that of the miRNAs•Changes in ceRNAs are typically too small to influence miRNA-mediated repression
Denzler et al. investigate the stoichiometric relationship of a microRNA and its target sites in primary cells. They find that microRNA target sites exceed microRNA copy numbers and that changes in the number of target sites that occur across physiological and disease conditions are typically too small to detectibly influence microRNA activity.
MicroRNAs (miRNAs) pair to sites in mRNAs to direct the degradation of these RNA transcripts. Conversely, certain RNA transcripts can direct the degradation of particular miRNAs. This target-directed ...miRNA degradation (TDMD) requires the ZSWIM8 E3 ubiquitin ligase. Here, we report the function of ZSWIM8 in the mouse embryo.
embryos were smaller than their littermates and died near the time of birth. This highly penetrant perinatal lethality was apparently caused by a lung sacculation defect attributed to failed maturation of alveolar epithelial cells. Some mutant individuals also had heart ventricular septal defects. These developmental abnormalities were accompanied by aberrant accumulation of more than 50 miRNAs observed across 12 tissues, which often led to enhanced repression of their mRNA targets. These ZSWIM8-sensitive miRNAs were preferentially produced from genomic miRNA clusters, and in some cases, ZSWIM8 caused a switch in the dominant strand or isoform that accumulated from a miRNA hairpin-observations suggesting that TDMD provides a mechanism to uncouple coproduced miRNAs from each other. Overall, our findings indicate that the regulatory influence of ZSWIM8, and presumably TDMD, in mammalian biology is widespread and consequential, and posit the existence of many yet-unidentified transcripts that trigger miRNA degradation.
D-cyclins represent components of cell cycle machinery. To test the efficacy of targeting D-cyclins in cancer treatment, we engineered mouse strains that allow acute and global ablation of individual ...D-cyclins in a living animal. Ubiquitous shutdown of cyclin D1 or inhibition of cyclin D-associated kinase activity in mice bearing ErbB2-driven mammary carcinomas triggered tumor cell senescence, without compromising the animals’ health. Ablation of cyclin D3 in mice bearing Notch1-driven T cell acute lymphoblastic leukemias (T-ALL) triggered tumor cell apoptosis. Such selective killing of leukemic cells can also be achieved by inhibiting cyclin D associated kinase activity in mouse and human T-ALL models. Inhibition of cyclin D-kinase activity represents a highly-selective anticancer strategy that specifically targets cancer cells without significantly affecting normal tissues.
► Acute and global shutdown of cyclins D1 or D3 does not compromise animals’ health ► Acute cyclin D1 ablation in mice with breast cancers causes tumor cell senescence ► Ablation of cyclin D3 selectively kills leukemic cells in a mouse model of T-ALL ► Inhibition of cyclin D-CDK kinase causes apoptosis of human T-ALL cells in vivo
D-type cyclins (D1, D2, and D3) are components of the mammalian core cell-cycle machinery and function to drive cell proliferation. Here, we report that D-cyclins perform a rate-limiting ...antiapoptotic function in vivo. We found that acute shutdown of all three D-cyclins in bone marrow of adult mice resulted in massive apoptosis of all hematopoietic cell types. We demonstrate that adult hematopoietic stem cells are particularly dependent on D-cyclins for survival and that they are especially sensitive to cyclin D loss. Surprisingly, we found that the antiapoptotic function of D-cyclins also operates in quiescent hematopoietic stem and progenitor cells. Our analyses revealed that D-cyclins repress the expression of the death receptor Fas and its ligand, FasL. Acute ablation of D-cyclins upregulated these proapoptotic genes and led to Fas- and caspase 8-dependent apoptosis. These results reveal an unexpected function of cell-cycle proteins in controlling apoptosis in normal cell homeostasis.
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•D-cyclins perform a rate-limiting antiapoptotic function in vivo•D-cyclins control the levels of Fas and Fas ligand via E2F1 and repress cell death•Adult hematopoietic stem cells are particularly dependent on D-cyclins for survival•D-cyclins play an antiapoptotic function also in quiescent stem/progenitor cells
D-type cyclins belong to the core cell-cycle machinery and function to drive cell proliferation. Choi et al. report that D-cyclins also perform a major antiapoptotic function in hematopoietic cells. Acute shutdown of D-cyclins in bone marrow triggered massive apoptosis, and hematopoietic stem cells were particularly sensitive to cyclin D ablation.
The accumulation of large boulders related to waves generated by either tsunamis or extreme storm events have been observed in different areas of the Mediterranean Sea. Along the eastern low-lying ...rocky coasts of Malta, five sites with large boulder deposits have been investigated, measured and mapped. These boulders have been detached and moved from the nearshore and the lowest parts of the coast by sea wave action. In the Sicily–Malta channel, heavy storms are common and originate from the NE and NW winds. Conversely, few tsunamis have been recorded in historical documents to have reached the Maltese archipelago. We present a multi-disciplinary study, which aims to define the characteristics of these boulder accumulations, in order to assess the coastal geo-hazard implications triggered by the sheer ability of extreme waves to detach and move large rocky blocks inland. The wave heights required to transport 77 coastal boulders were calculated using various hydrodynamic equations. Particular attention was given to the quantification of the input parameters required in the workings of these equations, such as size, density and distance from the coast. In addition, accelerator mass spectrometry (AMS) 14C ages were determined from selected samples of marine organisms encrusted on some of the coastal boulders. The combination of the results obtained both by the hydrodynamic equations, which provided values comparable with those observed and measured during the storms, and radiocarbon dating suggests that the majority of the boulders have been detached and moved by intense storm waves. These boulders testify to the existence of a real hazard for the coasts of Malta, i.e. that of very high storm waves, which, during exceptional storms, are able to detach large blocks of volumes exceeding 10 m3 from the coastal edge and the nearshore bottom, and also to transport them inland. Nevertheless, the occurrence of one or more tsunami events cannot be ruled out, since radiocarbon dating of some marine organisms did reveal ages which may be related to historically known tsunamis in the Mediterranean region, such as the ones in AD 963, 1329, 1693 and 1743.
ABSTRACT
We analyse the chemical properties of three z∼ 8 galaxies behind the galaxy cluster SMACS J0723.3-7327, observed as part of the Early Release Observations programme of the James Webb Space ...Telescope. Exploiting O iiiλ4363 auroral line detections in NIRSpec spectra, we robustly apply the direct Te method for the very first time at such high redshift, measuring metallicities ranging from extremely metal poor (12 + log(O/H)≈ 7) to about one-third solar. We also discuss the excitation properties of these sources, and compare them with local strong-line metallicity calibrations. We find that none of the considered diagnostics match simultaneously the observed relations between metallicity and strong-line ratios for the three sources, implying that a proper re-assessment of the calibrations may be needed at these redshifts. On the mass–metallicity plane, the two galaxies at z ∼ 7.6 ($\rm log(M_*/M_{\odot }) = 8.1, 8.7$) have metallicities that are consistent with the extrapolation of the mass–metallicity relation at z∼2–3, while the least massive galaxy at z ∼ 8.5 ($\rm log(M_*/M_{\odot }) = 7.8$) shows instead a significantly lower metallicity. The three galaxies show different level of offset relative to the Fundamental Metallicity Relation, with two of them (at z∼ 7.6) being marginally consistent, while the z∼ 8.5 source deviating significantly, being probably far from the smooth equilibrium between gas flows, star formation, and metal enrichment in place at later epochs.
Summary Background The absence of trial data comparing robot-assisted laparoscopic prostatectomy and open radical retropubic prostatectomy is a crucial knowledge gap in uro-oncology. We aimed to ...compare these two approaches in terms of functional and oncological outcomes and report the early postoperative outcomes at 12 weeks. Method In this randomised controlled phase 3 study, men who had newly diagnosed clinically localised prostate cancer and who had chosen surgery as their treatment approach, were able to read and speak English, had no previous history of head injury, dementia, or psychiatric illness or no other concurrent cancer, had an estimated life expectancy of 10 years or more, and were aged between 35 years and 70 years were eligible and recruited from the Royal Brisbane and Women's Hospital (Brisbane, QLD). Participants were randomly assigned (1:1) to receive either robot-assisted laparoscopic prostatectomy or radical retropubic prostatectomy. Randomisation was computer generated and occurred in blocks of ten. This was an open trial; however, study investigators involved in data analysis were masked to each patient's condition. Further, a masked central pathologist reviewed the biopsy and radical prostatectomy specimens. Primary outcomes were urinary function (urinary domain of EPIC) and sexual function (sexual domain of EPIC and IIEF) at 6 weeks, 12 weeks, and 24 months and oncological outcome (positive surgical margin status and biochemical and imaging evidence of progression at 24 months). The trial was powered to assess health-related and domain-specific quality of life outcomes over 24 months. We report here the early outcomes at 6 weeks and 12 weeks. The per-protocol populations were included in the primary and safety analyses. This trial was registered with the Australian New Zealand Clinical Trials Registry (ANZCTR), number ACTRN12611000661976. Findings Between Aug 23, 2010, and Nov 25, 2014, 326 men were enrolled, of whom 163 were randomly assigned to radical retropubic prostatectomy and 163 to robot-assisted laparoscopic prostatectomy. 18 withdrew (12 assigned to radical retropubic prostatectomy and six assigned to robot-assisted laparoscopic prostatectomy); thus, 151 in the radical retropubic prostatectomy group proceeded to surgery and 157 in the robot-assisted laparoscopic prostatectomy group. 121 assigned to radical retropubic prostatectomy completed the 12 week questionnaire versus 131 assigned to robot-assisted laparoscopic prostatectomy. Urinary function scores did not differ significantly between the radical retropubic prostatectomy group and robot-assisted laparoscopic prostatectomy group at 6 weeks post-surgery (74·50 vs 71·10; p=0·09) or 12 weeks post-surgery (83·80 vs 82·50; p=0·48). Sexual function scores did not differ significantly between the radical retropubic prostatectomy group and robot-assisted laparoscopic prostatectomy group at 6 weeks post-surgery (30·70 vs 32·70; p=0·45) or 12 weeks post-surgery (35·00 vs 38·90; p=0·18). Equivalence testing on the difference between the proportion of positive surgical margins between the two groups (15 10% in the radical retropubic prostatectomy group vs 23 15% in the robot-assisted laparoscopic prostatectomy group) showed that equality between the two techniques could not be established based on a 90% CI with a Δ of 10%. However, a superiority test showed that the two proportions were not significantly different (p=0·21). 14 patients (9%) in the radical retropubic prostatectomy group versus six (4%) in the robot-assisted laparoscopic prostatectomy group had postoperative complications (p=0·052). 12 (8%) men receiving radical retropubic prostatectomy and three (2%) men receiving robot-assisted laparoscopic prostatectomy experienced intraoperative adverse events. Interpretation These two techniques yield similar functional outcomes at 12 weeks. Longer term follow-up is needed. In the interim, we encourage patients to choose an experienced surgeon they trust and with whom they have rapport, rather than a specific surgical approach. Funding Cancer Council Queensland.
Female primates signal impending ovulation with a suite of sexual signals. Studies of these signals have focussed on visual, and to a lesser extent, acoustic signals, neglecting olfactory signals. We ...aimed to investigate the information content of female olfactory signals in captive olive baboons (Papio anubis) and relate these to the female fertile period. We studied eight adult females living in four groups at the CNRS Station de Primatologie, Rousset-sur-Arc, France. We used vaginal cytology to detect ovulation. We investigated the volatile component of odour signals using solid-phase microextraction and gas chromatography-mass spectrometry. We found a total of 74 volatile compounds, of which we tentatively identified 25, including several ketones, alcohols, aldehydes, terpenes, volatile fatty acids and hydrocarbons that have been identified in odour profiles of other primates. Our results show that vaginal odour intensity differs with sexual cycle stage suggesting that odour might play a role in signalling female baboon fertility. We found differences in vaginal odour between females living in all-female and in mixed sex groups but we could not distinguish the effects of group composition, female age and identity. This study of olfactory signalling improves our understanding of how female primates advertise their sexual receptivity.
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