Detection of snRNP assembly intermediates in Cajal bodies by fluorescence resonance energy transfer

Spliceosomal small nuclear ribonucleoprotein particles (snRNPs) are required for pre-mRNA splicing throughout the nucleoplasm, yet snRNPs also concentrate in Cajal bodies (CBs). To address a proposed role of CBs in snRNP assembly, we have used fluorescence resonance energy transfer (FRET) microscopy to investigate the subnuclear distribution of specific snRNP intermediates. Two distinct complexes containing the protein SART3 (p110), required for U4/U6 snRNP assembly, were localized: SART3•U6 snRNP and SART3•U4/U6 snRNP. These complexes segregated to different nuclear compartments, with SART3•U6 snRNPs exclusively in the nucleoplasm and SART3•U4/U6 snRNPs preferentially in CBs. Mutant cells lacking the CB-specific protein coilin and consequently lacking CBs exhibited increased nucleoplasmic levels of SART3•U4/U6 snRNP complexes. Reconstitution of CBs in these cells by expression of exogenous coilin restored accumulation of SART3•U4/U6 snRNP in CBs. Thus, while some U4/U6 snRNP assembly can occur in the nucleoplasm, these data provide evidence that SART3•U6 snRNPs form in the nucleoplasm and translocate to CBs where U4/U6 snRNP assembly occurs

Granite microporosity changes due to fracturing and alteration: secondary mineral phases as proxies for porosity and permeability estimation

Several alteration facies of fractured Lipnice granite are studied in detail on borehole samples by means of mercury intrusion porosimetry, polarized and fluorescent light microscopy, and microprobe chemical analyses. The goal is to describe the granite void space geometry in the vicinity of fractures with alteration halos and to link specific geometries with simply detectable parameters to facilitate quick estimation of porosity and permeability based on, for example, drill cuttings. The core of the study is the results of porosity and throat size distribution analyses on 21 specimens representing unique combinations of fracture-related structures within six different alteration facies basically differing in secondary phyllosilicate chemistry and porosity structure. Based on a simple model to calculate permeability from the measured porosities and throat size distributions, the difference in permeability between the fresh granite and the most fractured and altered granite is 5 orders of magnitude. Our observations suggest that the porosity, the size of connections and the proportion of crack porosity increase with fracture density, while precipitation of iron-rich infills as well as of fine-grained secondary phyllosilicates acts in the opposite way. Different styles and intensities of such end-member agents shape the final void space geometry and imply various combinations of storage, transport and retardation capacity for specific structures. This study also shows the possibility to use standard mercury intrusion porosimetry with advanced experimental settings and data treatment to distinguish important differences in void space geometry within a span of a few percent of porosity.</p

Assessment of spectral UV radiation at Marambio Base, Antarctic Peninsula

This study aims to assess the dependence of spectral UV radiation on different atmospheric and terrestrial factors, including solar zenith angle, ozone, and cloud cover, in the southern polar environment. For this purpose, 23 260 spectra (300–363 nm), obtained by the B199 Mk-III Brewer spectrophotometer at Marambio Base, Antarctic Peninsula region, over the period 2010–2020, were studied. A neural network model was developed to investigate the effects of the explanatory variables at 127 wavelengths in the interval 300–363 nm, with a 0.5 nm sampling interval. Solar zenith angle (SZA) proved to be the most important parameter, followed by cloud cover, total ozone column (TOC), and surface albedo. The relative SZA effect is greatest at the shortest wavelengths, where a 1∘ decrease in SZA results in a 6 %–18 % increase in UV irradiance (305 nm). TOC particularly affects the short wavelengths below approximately 320–325 nm, when for example at 305 nm a 10 DU decrease in TOC causes a 7 %–13 % increase in UV irradiance. The large-scale ozone holes (e.g., in 2011–2012, 2014–2015, 2018–2019) caused the spectral UV irradiance at very short wavelengths to peak in spring, whereas in other seasons (e.g., 2010–2011, 2012–2013), the maxima at all wavelengths were recorded in summer (November to January). Absorption of UV radiance by the ozone also affected the temporal distribution of very high spectral UV irradiances (i.e., highest 10 % of the distribution), when at 305 nm they were observed both in spring and summer months, and at 340 nm they occurred mostly in summer. The effect of cloud cover was strongest near the fully cloudy sky and in the summer months, when the Antarctic clouds tend to be thickest.</p

Histone Deacetylase Activity Modulates Alternative Splicing

There is increasing evidence to suggest that splicing decisions are largely made when the nascent RNA is still associated with chromatin. Here we demonstrate that activity of histone deacetylases (HDACs) influences splice site selection. Using splicing-sensitive microarrays, we identified ∼700 genes whose splicing was altered after HDAC inhibition. We provided evidence that HDAC inhibition induced histone H4 acetylation and increased RNA Polymerase II (Pol II) processivity along an alternatively spliced element. In addition, HDAC inhibition reduced co-transcriptional association of the splicing regulator SRp40 with the target fibronectin exon. We further showed that the depletion of HDAC1 had similar effect on fibronectin alternative splicing as global HDAC inhibition. Importantly, this effect was reversed upon expression of mouse HDAC1 but not a catalytically inactive mutant. These results provide a molecular insight into a complex modulation of splicing by HDACs and chromatin modifications

The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells

GFP-tagged snRNP components reveal the dynamics and rate for spliceosome assembly in vivo