The special Sm core structure of the U7 snRNP: far-reaching significance of a small nuclear ribonucleoprotein

The polypeptide composition of the U7 small nuclear ribonucleoprotein (snRNP) involved in histone messenger RNA (mRNA) 3' end formation has recently been elucidated. In contrast to spliceosomal snRNPs, which contain a ring-shaped assembly of seven so-called Sm proteins, in the U7 snRNP the Sm proteins D1 and D2 are replaced by U7-specific Sm-like proteins, Lsm10 and Lsm11. This polypeptide composition and the unusual structure of Lsm11, which plays a role in histone RNA processing, represent new themes in the biology of Sm/Lsm proteins. Moreover this structure has important consequences for snRNP assembly that is mediated by two complexes containing the PRMT5 methyltransferase and the SMN (survival of motor neurons) protein, respectively. Finally, the ability to alter this polypeptide composition by a small mutation in U7 snRNA forms the basis for using modified U7 snRNA derivatives to alter specific pre-mRNA splicing events, thereby opening up a new way for antisense gene therapy.

Unique Sm core structure of U7 snRNPs: assembly by a specialized SMN complex and the role of a new component, Lsm11, in histone RNA processing.

A set of seven Sm proteins assemble on the Sm-binding site of spliceosomal U snRNAs to form the ring-shaped Sm core. The U7 snRNP involved in histone RNA 3' processing contains a structurally similar but biochemically unique Sm core in which two of these proteins, Sm D1 and D2, are replaced by Lsm10 and by another as yet unknown component. Here we characterize this factor, termed Lsm11, as a novel Sm-like protein with apparently two distinct functions. In vitro studies suggest that its long N-terminal part mediates an important step in histone mRNA 3'-end cleavage, most likely by recruiting a zinc finger protein previously identified as a processing factor. In contrast, the C-terminal part, which comprises two Sm motifs interrupted by an unusually long spacer, is sufficient to assemble with U7, but not U1, snRNA. Assembly of this U7-specific Sm core depends on the noncanonical Sm-binding site of U7 snRNA. Moreover, it is facilitated by a specialized SMN complex that contains Lsm10 and Lsm11 but lacks Sm D1/D2. Thus, the U7-specific Lsm11 protein not only specifies the assembly of the U7 Sm core but also fulfills an important role in U7 snRNP-mediated histone mRNA processing.

3' end processing of mouse histone pre-mRNA: evidence for additional base-pairing between U7 snRNA and pre-mRNA.

We have analysed the extent of base-pairing interactions between spacer sequences of histone pre-mRNA and U7 snRNA present in the trans-acting U7 snRNP and their importance for histone RNA 3' end processing in vitro. For the efficiently processed mouse H4-12 gene, a computer analysis revealed that additional base pairs could be formed with U7 RNA outside of the previously recognised spacer element (stem II). One complementarity (stem III) is located more 3' and involves nucleotides from the very 5' end of U7 RNA. The other, more 5' located complementarity (stem I) involves nucleotides of the Sm binding site of U7 RNA, a part known to interact with snRNP structural proteins. These potential stem structures are separated from each other by short internal loops of unpaired nucleotides. Mutational analyses of the pre-mRNA indicate that stems II and III are equally important for interaction with the U7 snRNP and for processing, whereas mutations in stem I have moderate effects on processing efficiency, but do not impair complex formation with the U7 snRNP. Thus nucleotides near the processing site may be important for processing, but do not contribute to the assembly of an active complex by forming a stem I structure. The importance of stem III was confirmed by the ability of a complementary mutation in U7 RNA to suppress a stem III mutation in a complementation assay using Xenopus laevis oocytes. The main role of the factor(s) binding to the upstream hairpin loop is to stabilise the U7-pre-mRNA complex. This was shown by either stabilising (by mutation) or destabilising (by increased temperature) the U7-pre-mRNA base-pairing under conditions where hairpin factor binding was either allowed or prevented (by mutation or competition). The hairpin dependence of processing was found to be inversely related to the strength of the U7-pre-mRNA interaction.

The low abundance of U7 snRNA is partly determined by its Sm binding site.

In transient expression studies after DNA transfection of HeLa cells, the mouse U7 gene produces only approximately 30% of the RNA produced by a mouse U1b gene. This difference persists even when the transfected genes have all their 5' and 3' flanking sequences exchanged suggesting a post-transcriptional effect. When the special U7 Sm binding site is mutated to a consensus derived from the major snRNAs (Sm-opt), the U7 RNA level increases 4- to 5-fold, whereas no RNA is detected from a U7 gene with a non-functional Sm binding site (Sm-mut). Moreover, U1b genes with the U7 Sm binding site yield reduced RNA levels. The Sm-opt site also alters the cellular behaviour of the corresponding U7 snRNA. It accumulates to a higher level in the nucleus than wild type U7 RNA, and is better immunoprecipitable with anti-Sm antibodies. Injection experiments in Xenopus oocytes indicate that the U7 genes with either Sm-opt or Sm-mut sites produce similar amounts of RNA as wild type U7, but that they differ in opposing ways in the processing of precursors to mature size U7 snRNA and in nuclear accumulation. However, in reconstitution experiments using Xenopus oocytes, we show that U7 Sm-opt RNA, despite its efficient nuclear accumulation, is not active in 3' processing of histone pre-mRNA, whereas wild type U7 RNA is assembled into functional snRNPs, which correctly process histone pre-mRNA substrate. This suggests a functional importance of the special U7 Sm sequence.

Fragm. lat. I 10 - Breviarium (Dom. V, VI, XXI-XXIII p.Pent.; Lesungen aus I Sm und Prv)

Trägerband: Inc. qu. 1231; Ms. Praed. 116; Vorbesitzer: Dominikanerkloster Frankfurt am Main

Facies investigations on sediment core CRP-3 from the Ross Sea, Antarctica

The Cenozoic Victoria Land Basin (VLB) stratigraphic section penetrated by CRP-3 is mostly of Early Oligocene age. It contains an array of lithofacies comprising fine-grained mudrocks, interlaminated and interbedded mudrocks/sandstones, mud-rich and mud-poor sandstones, conglomerates and diamctites that are together interpreted as the products of shallow marine to possibly non-marine environments of deposition, affected by the periodic advance and retreat of tidewater glaciers. This lithofacies assemblage can be readily rationalised using the facies scheme designed originally for CRP-2/2A, and published previously. The uppermost 330 metres below sea floor (mbsf) shows a cyclical arrangement of lithofacies also similar to that recognised throughout CRP-2/2A, and interpreted to reflect cyclical variations in relative sea-level driven by ice volume fluctuations ('Motif A'). Between 330 and 480 mbsf, a series of less clearly cyclical units, generally fining-upward but nonetheless incorporating a significant subset of the facies assemblage, has been identified and noted in the Initial Report as 'Motif B' Below 480 mbsf, the section is arranged into a repetitive succession of fining-upward units, each of which comprises dolerite clast conglomerate at the base passing upward into relatively thick intervals of sandstones. The cycles present down 480 mbsf are defined as sequences, each interpreted to record cyclical variation of relative sea-level. The thickness distribution of sequences in CRP-3 provides some insights into the geological variables controlling sediment accumulation in the Early Oligocene section. The uppermost part of the section in CRP-3 comprises two or three thick, complete sequences that show a broadly symmetrical arrangement of lithofacies (similar to Sequences 9-11 in CRP-2/2A). This suggests a period of relatively rapid tectonic subsidence, which allowed preservation of the complete facies cycle. Below Sequence 3, however, is a considerable interval of thin, incomplete and erosionally truncated sequences (4-23), which incorporates both the remainder of Motif A sequences and all Motif B sequences recognised. The thinner and more truncated sequences suggest sediment accumulation under conditions of reduced accommodation, and given the lack of evidence for glacial conditions (see Powell et al., this volume) tends to argue for a period of reduced tectonic subsidence. The section below 480 mbsf consists of a series of fining-upward, conglomerate to sandstone intervals which cannot be readily interpreted in terms of relative sea-level change. A relatively mudrock-rich interval above the basal conglomerate/breccia (782-762 mbsf) may record initial flooding of the basin during early rift subsidence. The lithostratigraphy summarised above has been linked to seismic reflection data using depth conversion techniques (Henrys et al., this volume). The three uppermost reflectors ('o', 'p' and 'q') correlate to the package of thick sequences 1-3, and several deeper reflectors can also be correlated to sequence boundaries. The package of thick Sequences 1-3 shows a sheet-like cross-sectional geometry on seismic reflection lines, unlike the similar package recognised in CRP-2/2A.

Rb, Sr, Sm and Nd concentrations and isotopic composition of basalts from DSDP Leg 74 (Table 1)

Basement intersected in DSDP holes 525A, 528 and 527 on the Walvis Ridge consists of submarine basalt flows and pillows with minor intercalated sediments. These holes are situated on the crest and mid and lower northwest flank of a NNW-SSE-trending ridge block which would have closely paralleled the paleo mid-ocean ridge (Rabinowitz and LaBrecque, 1979 doi:10.1029/JB084iB11p05973, Moore et al. (1983 doi:10.1130/0016-7606(1983)94<907:TWRTDS>2.0.CO;2). The basalts were erupted approximately 70 m.y. ago, an age equivalent to that of immediately adjacent oceanic crust in the Angola Basin and coraistent with formation at the paleo mid-ocean ridge (Moore et al., 1983). The basalt types vary from aphyric quartz tholeiites on the ridge crest to highly plagioclase phyric olivine tholeiites on the ridge flank. These show systematic differences in incompatible trace element and isotopic composition. Many element and isotope ratio pairs form systematic trends with the ridge crest basalts at one end and the highly phyric ridge flank basalts at the other. The low 143Nd/144Nd (0.51238), 206Pb/204Pb (17.54), 207Pb/204Pb (15.47), 208Pb/204Pb (38.14) and high 87Sr/86Sr (0.70512) ratios of the ridge crest basalts suggest derivation from an old Nd/Sm-, Rb/Sr- and Pb/U-enriched mantle source. This isotopic signature is similar to that of alkaline basalts on Tristan da Cunha but offset to significantly lower Nd and Pb isotopic ratios. The isotopic ratio trends may be extrapolated beyond the ridge flank basalts with higher 143Nd/144Nd (0.51270), 206Pb/204Pb (18.32), 207Pb/204Pb (15.52), 208Pb/204Pb (38.77) and lower 87Sr/86Sr (0.70417) ratios in the direction of increasingly Nd/Sm-, Rb/Sr- and Pb/U-depleted source compositions. These isotopic correlations are equally consistent with mixing of depleted and enriched end member melts or partial melting of an inhomogeneous, variably enriched mantle source. However, observed Zr-Ba-Nb-Y interelement relationships are inconsistent with any simple two-component model of magma mixing, as might result from the rise of a lower mantle plume through the upper mantle. Incompatible element and Pb isotopic systematics also preclude extensive involvement of depleted (N-type) MORB material or its mantle sources. In our preferred petrogenetic model the Walvis Ridge basalts were derived by partial melting of mantle similar to an enriched (E-type) MORB source which had become heterogeneous on a small scale due to the introduction of small-volume melts and metasomatic fluids.

(Table 1) Isotopic composition of Sm and Nd in mantle restites of the Central Atlantic and in associated plutonic and volcanic rocks

Ultrabasic rock samples collected from two areas of the crustal zone of the Mid-Atlantic Ridge (MAR): (1) 13-17°N (near the intersection of the ridge axis with the 15°20'N prime fracture zone), and (2) 33°40'N prime (the western intersection of the MAR crest with the Heis fracture zone) were objects of this study. Samples of peridotite and of plutonic and volcanic rocks associated with it were used to measure their Sm/Nd, 143Nd/144Nd, and 147Sm/144Nd ratios, which allowed to test time and genetic relationships between evolution of mantle material under the ridge crest and products of its magmatic activity. Results of this work proved ubiquitous discrepancy between melting degree values of extremely depleted mantle peridotites in the MAR area between 14°N and 16°N, obtained using petrologic and geochemical methods. This discrepancy suggests large-scale interaction between mantle material and magmatic melts and fluids enriched in incompatible elements or fluids. The results obtained suggest that repeated melting of the mantle under the axial MAR zone is an universal characteristic of magmatism in low-velocity spreading centers. The results of this study also proved the crestal MAR zone in the Central Atlantic region show distinct indications of isotope-geochemical segmentation of the mantle. It is suggested that the geochemically anomalous MAR mantle peridotite in the zone of the MAR intersection with the 15°20'N prime fracture zone can be interpreted as fragments of mantle substrate, foreign for the Atlantic mantle north of the equator.

Sm-Nd isotopic investigation for rocks of Prince Charles Mountains, East Antarctica

The Prince Charles Mountains have been subject to extensive geological and geophysical investigations by former Soviet, Russian and Australian scientists from the early 1970s. In this paper we summarise, and review available geological and isotopic data, and report results of new isotopic studies (Sm-Nd, Pb-Pb, and U-Pb SHRIMP analyses); field geological data obtained during the PCMEGA 2002/2003 are utilised. The structure of the region is described in terms of four tectonic terranes. Those include Archaean Ruker, Palaeoproterozoic Lambert, Mesoproterozoic Fisher, and Meso- to Neoproterozoic Beaver Terranes. Pan-African activities (granite emplacement and probably tectonics) in the Lambert Terrane are reported. We present a summary of the composition of these terranes, discuss their origin and relationships. We also outline the most striking geological features, and problems, and try to draw attention to those rocks and regional geological features which are important in understanding the composition and evolution of the PCM and might suggest targets for further investigations.