Identification of a Novel 81-kDa Component of the Xenopus Origin Recognition Complex

The Xenopus origin recognition complex is essential for chromosomal DNA replication in cell-free extracts. We have immunopurified the Xenopus origin recognition complex with anti-Xorc2 antibodies and analyzed its composition and properties. Xorc2 (p63) is specifically associated with Xorc1 (p115) and up to four additional polypeptides (p81, p78, p45, and p40). The cDNA encoding p81 is highly homologous to various expressed sequence tags from humans and mice encoding a protein of previously unknown function. Immunodepletion of p81 from Xenopus egg extracts, which also results in the removal of Xorc2, completely abolishes chromosomal DNA replication. Thus, p81 appears to play a crucial role at S phase in higher eukaryotes

137CS gamma-ray detection at Summit, Greenland

Global fall-out from atmospheric testing of thermonuclear weapons produced horizon markers corresponding to the initiation of testing in 1953 and the maximum fall-out in 1963. The radioactive isotope 137Cs associated with these events has a half-life of 30.2 years. Therefore, with the appropriate radiation detectors, this fall-out can be used as a long-term temporal indicator in glaciers and snowpack. A prototype γ-ray detector system was successfully tested and was used to make in-situ measurements of the 137Cs marker in a borehole at Summit, Greenland. The system consisted of a 7.6 cm by 7.6 cm NaI (Tl) scintillation crystal/photomultiplier detector, commercial pre-amplifier, amplifier and power supplies, and a microcomputer-based pulse-height analyzer. The measurements were made in boreholes of 25.4 cm and 12.7 cm diameter to depths of 22 m. Based on the results reported here, the γ-ray detection technique promises to be a powerful way to locate quickly horizon markers in the field. -Author

The Rad9-Hus1-Rad1 Checkpoint Clamp Regulates Interaction of TopBP1 with ATR

TopBP1 serves as an activator of the ATR-ATRIP complex in response to the presence of incompletely replicated or damaged DNA. This process involves binding of ATR to the ATR-activating domain of TopBP1, which is located between BRCT domains VI and VII. TopBP1 displays increased binding to ATR-ATRIP in Xenopus egg extracts containing checkpoint-inducing DNA templates. We show that an N-terminal region of TopBP1 containing BRCT repeats I-II is essential for this checkpoint-stimulated binding of TopBP1 to ATR-ATRIP. The BRCT I-II region of TopBP1 also binds specifically to the Rad9-Hus1-Rad1 (9-1-1) complex in Xenopus egg extracts. This binding occurs via the C-terminal domain of Rad9 and depends upon phosphorylation of its Ser-373 residue. Egg extracts containing either a mutant of TopBP1 lacking the BRCT I-II repeats or a mutant of Rad9 with an alanine substitution at Ser-373 are defective in checkpoint regulation. Furthermore, an isolated C-terminal fragment from Rad9 is an effective inhibitor of checkpoint signaling in egg extracts. These findings suggest that interaction of the 9-1-1 complex with the BRCT I-II region of TopBP1 is necessary for binding of ATR-ATRIP to the ATR-activating domain of TopBP1 and the ensuing activation of ATR

Claspin and the Activated Form of ATR-ATRIP Collaborate in the Activation of Chk1

Claspin is necessary for the ATR-dependent activation of Chk1 in Xenopus egg extracts containing incompletely replicated DNA. ATR possesses a regulatory partner called ATRIP. We have studied the respective roles of ATR-ATRIP and Claspin in the activation of Chk1. ATR-ATRIP bound well to various DNA templates in Xenopus egg extracts. ATR-ATRIP bound to a single-stranded DNA template was weakly active. By contrast, the ATR-ATRIP complex on a DNA template containing both single- and double-stranded regions displayed a large increase in kinase activity. This observation suggests that ATR-ATRIP normally undergoes activation upon association with specific nucleic acid structures at DNA replication forks. Without Claspin, activated ATR-ATRIP phosphorylated Chk1 weakly in a cell-free reaction. The addition of Claspin to this reaction strongly stimulated the phosphorylation of Chk1 by ATR-ATRIP. Claspin also induced significant autophosphorylation of Chk1 in the absence of ATR-ATRIP. Taken together, these results indicate that the checkpoint-dependent phosphorylation of Chk1 is a multistep process involving activation of the ATR-ATRIP complex at replication forks and presentation of Chk1 to this complex by Claspin

Beyond Nationalism? The Anti-Austerity Social Movement in Ireland:Between Domestic Constraints and Lessons from Abroad

The European financial crisis has inspired a wave of social activism, challenging established party politics. In Ireland, a large social protest movement produced a Right2Change political campaign in 2015 that confronted the right-wing consensus in Irish politics. Some activists sought to emulate and learn from the example of new parties like Spain’s Podemos. Yet, to date, the traditional party structure remains intact, and hopes of emulating the success of activists elsewhere remain muted. At the same time, anti-austerity activism in Ireland has seen a conscious attempt to engage in intense dialogue with the pan-European experience. Irish activists have looked to Europe before for inspiration and a sharing of experiences; but the most recent attempt to create a Europeanised public space in response to the perceived ‘blockage’ in the Irish party system is something fairly innovative. This article, based in part on ten semi-structured interviews with politicians and social movement activists, considers the achievements and failings of the Irish anti-austerity movement to date.</p

Cell Cycle Control by Xenopus p28^(Kix1) a Developmentally Regulated Inhibitor of Cyclin-dependent Kinases

We have isolated Xenopus p28^(Kix1), a member of the p21^(CIP1)/p27^(KIP1)/p57^(KIP2) family of cyclin-dependent kinase (Cdk) inhibitors. Members of this family negatively regulate cell cycle progression in mammalian cells by inhibiting the activities of Cdks. p28 shows significant sequence homology with p21, p27, and p57 in its N-terminal region, where the Cdk inhibition domain is known to reside. In contrast, the C-terminal domain of p28 is distinct from that of p21, p27, and p57. In co-immunoprecipitation experiments, p28 was found to be associated with Cdk2, cyclin E, and cyclin A, but not the Cdc2/cyclin B complex in Xenopus egg extracts. Xenopus p28 associates with the proliferating cell nuclear antigen, but with a substantially lower affinity than human p21. In kinase assays with recombinant Cdks, p28 inhibits pre-activated Cdk2/cyclin E and Cdk2/cyclin A, but not Cdc2/cyclin B. However, at high concentrations, p28 does prevent the activation of Cdc2/cyclin B by the Cdk-activating kinase. Consistent with the role of p28 as a Cdk inhibitor, recombinant p28 elicits an inhibition of both DNA replication and mitosis upon addition to egg extracts, indicating that it can regulate multiple cell cycle transitions. The level of p28 protein shows a dramatic developmental profile: it is low in Xenopus oocytes, eggs, and embryos up to stage 11, but increases approximately ~ 100-fold between stages 12 and 13, and remains high thereafter. The induction of p28 expression temporally coincides with late gastrulation. Thus, although p28 may play only a limited role during the early embryonic cleavages, it may function later in development to establish a somatic type of cell cycle. Taken together, our results indicate that Xenopus p28 is a new member of the p21/p27/p57 class of Cdk inhibitors, and that it may play a role in developmental processes

Children’s participation rights in early childhood education and care: the case of early literacy learning and pedagogy

This position article argues that educators’ knowledge of young children’s perspectives on aspects of early learning, including literacy learning, and subsequent interpretations of the ways that these perspectives can inform and shape pedagogy are key to promoting children’s participation rights in early childhood education and care. Drawing on ideas such as guided participation and Bruner’s notion of a pedagogy of mutuality, it is argued that pedagogy, as it is now understood, implies that children’s participation is central to the task of teaching. It is also argued that explicit articulation of the concept of joint participation in relation to literacy curricula and pedagogy in early childhood is warranted. The latter part of the article raises the issue of the diversity of children’s perspectives of literacy learning and pedagogy. There follows a discussion of some of the issues related to working with young children’s perspectives on literacy

Early childhood mathematics teaching: challenges, difficulties and priorities of teachers of young children in primary schools in Ireland

Issues of pedagogy are critical in all aspects of early childhood education. Early childhood mathematics is no exception. There is now a great deal of guidance available to teachers in terms of high-quality early childhood mathematics teaching. Consequently, the characteristics of high-quality early childhood mathematics education are clearly identifiable. Issues such as building on young children’s prior-to-school knowledge; engaging children in general mathematical processes; and assessing and documenting children’s learning are some of the key aspects of high-quality early childhood mathematics education. The extent to which teachers of four- and five-year-old children in primary schools in Ireland incorporate current pedagogical guidance in early childhood mathematics education was explored in 2007 in a nationally representative questionnaire survey of teachers of four- and five-year-old children attending primary schools. This paper presents some of the findings of the study in relation to teachers’ selfreported challenges, difficulties and priorities in teaching early childhood mathematics. Implications are drawn for professional development, curriculum guidance and educational policy