Die Eingliederung von Schwerbehinderten in das Arbeitsleben aus der Sicht von Unternehmen : Ergebnisse einer empirischen Untersuchung (Company estimation of the inte-gration of the severely disabled into working life : Results of an empirical study)

"The official figures of the unemployment statistics reveal the problematic situation on the labour market in black and white every time they are updated. Another development to be taken equally seriously stands out for a particular group in this army of unemployed people: unemployed severely disabled people. These people, too, are finding it increasingly difficult to obtain an adequate position. Besides their disability the loss of a job comes as an additional difficulty for them and is frequently the first step on the path to social isolation. Since the 1980s there has been a decline in the proportion of severely disabled people in the labour force as a whole in the Federal Republic of Germany - in spite of all the efforts made by labour administration and the main welfare institutions towards the integration of this group into the primary labour market. Against the background of this a project was introduced in Rheinland-Pfalz in 1994 which focuses on the (re)integration of severely disabled people into working life. In order to achieve this, those participating in the project pursue two different paths. Firstly in the companies, representatives of the disabled and works councils and secondly those responsible for personnel matters and/or personnel managers are contacted and are each offered special support services for the integration of severely disabled people. The effectiveness of these measures is additionally assessed by means of a scientific research parallel to this.What was of interest in the parallel research was first of all which area the integration efforts would affect. For this purpose a total of 4800 companies in four regions were asked to give an assessment of the integration. It is obvious that on the one hand the companies do not see anything in the existing qualifications which would constitute a serious obstacle to considering the recruitment of a disabled person. On the other hand it is clear that there is a considerable lack of information as to what support services and support opportunities are available for a company when opting to employ a disabled person." (Author's abstract, IAB-Doku) ((en))Schwerbehinderte, Arbeitsmarktchancen, Arbeitgeber - Einstellungen, betriebliche Integration, Arbeitssituation, Arbeitslosigkeitsbekämpfung, Qualifikationsanforderungen, Personalpolitik

Modified nucleotides at the 5′ end of human U2 snRNA are required for spliceosomal E-complex formation

U2 snRNA, a key player in nuclear pre-mRNA splicing, contains a 5′-terminal m(3)G cap and many internal modifications. The latter were shown in vertebrates to be generally required for U2 function in splicing, but precisely which residues are essential and their role in snRNP and/or spliceosome assembly is presently not clear. Here, we investigated the roles of individual modified nucleotides of HeLa U2 snRNA in pre-mRNA splicing, using a two-step in vitro reconstitution/complementation assay. We show that the three pseudouridines and five 2′O-methyl groups within the first 20 nucleotides of U2 snRNA, but not the m(3)G cap, are required for efficient pre-mRNA splicing. Individual pseudouridines were not essential, but had cumulative effects on U2 function. In contrast, four of five 2′O-methylations (at positions 1, 2, 12, and 19) were individually required for splicing. The in vitro assembly of 17S U2 snRNPs was not dependent on the presence of modified U2 residues. However, individual internal modifications were required for the formation of the ATP-independent early spliceosomal E complex. Our data strongly suggest that modifications within the first 20 nucleotides of U2 play an important role in facilitating the interaction of U2 with U1 snRNP and/or other factors within the E complex

Schulpädagogische Lehre braucht Unterrichtspraxis

Die Autoren stellen vor, wie wirkungsvoll ein früher Praxiseinsatz von Lehramtsstudierenden sein kann. Seminargruppen planen zusammen mit den Lehrpersonen und Hochschuldozierenden Unterricht, den sie dann gemeinsam in Schulklassen umsetzen und reflektieren. Vergleiche mit Seminargruppen, die keinen Schulbesuch absolvierten, zeigen, dass die Studierenden die praxisbezogenen Seminare in Bezug auf die Klarheit der Ausbildung, ihre eigene Motivation und ihre Planungskompetenz sehr viel positiver bewerten. (DIPF/Orig.

Schulpädagogische Lehre braucht Unterrichtspraxis

Die Autoren stellen vor, wie wirkungsvoll ein früher Praxiseinsatz von Lehramtsstudierenden sein kann. Seminargruppen planen zusammen mit den Lehrpersonen und Hochschuldozierenden Unterricht, den sie dann gemeinsam in Schulklassen umsetzen und reflektieren. Vergleiche mit Seminargruppen, die keinen Schulbesuch absolvierten, zeigen, dass die Studierenden die praxisbezogenen Seminare in Bezug auf die Klarheit der Ausbildung, ihre eigene Motivation und ihre Planungskompetenz sehr viel positiver bewerten. (DIPF/Orig.

Functional spliceosomal A complexes can be assembled in vitro in the absence of a penta-snRNP

Two different models currently exist for the assembly pathway of the spliceosome, namely, the traditional model, in which spliceosomal snRNPs associate in a stepwise, ordered manner with the pre-mRNA, and the holospliceosome model, in which all spliceosomal snRNPs preassemble into a penta-snRNP complex. Here we have tested whether the spliceosomal A complex, which contains solely U1 and U2 snRNPs bound to pre-mRNA, is a functional, bona fide assembly intermediate. Significantly, A complexes affinity-purified from nuclear extract depleted of U4/U6 snRNPs (and thus unable to form a penta-snRNP) supported pre-mRNA splicing in nuclear extract depleted of U2 snRNPs, whereas naked pre-mRNA did not. Mixing experiments with purified A complexes and naked pre-mRNA additionally confirmed that under these conditions, A complexes do not form de novo. Thus, our studies demonstrate that holospliceosome formation is not a prerequisite for generating catalytically active spliceosomes and that, at least in vitro, the U1 and U2 snRNPs can functionally associate with the pre-mRNA, prior to and independent of the tri-snRNP. The ability to isolate functional spliceosomal A complexes paves the way to study in detail subsequent spliceosome assembly steps using purified components

Analysis of site-specific protein–RNA cross-links in isolated RNP complexes, combining affinity selection and mass spectrometry

An important aspect of the assembly of RNPs, and in particular of spliceosomes, is the succession of proteins bound to any given site on the RNA. Protein–RNA cross-linking is a well-established technique for investigating this, but the identification of a cross-linked protein has so far relied upon the availability of antibodies for immunoprecipitation or Western blot studies. To facilitate identification of proteins independent of these techniques, site-specific protein–RNA cross-links were purified in a large scale, which were then used for mass spectrometry (MS). This approach was carried out by the use of a minimal pre-mRNA construct containing a single photoactivatable azidophenacyl group and an adjacent biotin-dT tag for affinity purification of the cross-linked product. To test the feasibility of the method, we purified cross-links to nucleotide 9 downstream of the 5′ splice site of pre-mRNA in the spliceosomal complexes A (“pre-spliceosome”) and H. By this method, we were able to identify several proteins by MS; the hnRNP proteins A2/B1 were cross-linked to the pre-mRNA in complex A, and FUSE 2/FBP (a homolog of the intronic splicing enhancer KSRP) was cross-linked in complex H

Spliceosomal U snRNP Core Assembly: Sm Proteins Assemble onto an Sm Site RNA Nonanucleotide in a Specific and Thermodynamically Stable Manner

The association of Sm proteins with U small nuclear RNA (snRNA) requires the single-stranded Sm site (PuAU(4–6)GPu) but also is influenced by nonconserved flanking RNA structural elements. Here we demonstrate that a nonameric Sm site RNA oligonucleotide sufficed for sequence-specific assembly of a minimal core ribonucleoprotein (RNP), which contained all seven Sm proteins. The minimal core RNP displayed several conserved biochemical features of native U snRNP core particles, including a similar morphology in electron micrographs. This minimal system allowed us to study in detail the RNA requirements for Sm protein-Sm site interactions as well as the kinetics of core RNP assembly. In addition to the uridine bases, the 2′ hydroxyl moieties were important for stable RNP formation, indicating that both the sugar backbone and the bases are intimately involved in RNA-protein interactions. Moreover, our data imply that an initial phase of core RNP assembly is mediated by a high affinity of the Sm proteins for the single-stranded uridine tract but that the presence of the conserved adenosine (PuAU…) is essential to commit the RNP particle to thermodynamic stability. Comparison of intact U4 and U5 snRNAs with the Sm site oligonucleotide in core RNP assembly revealed that the regions flanking the Sm site within the U snRNAs facilitate the kinetics of core RNP assembly by increasing the rate of Sm protein association and by decreasing the activation energy