Role of the ubiquitin-selective CDC-48/UFD-1/NPL-4 chaperone in DNA replication

Faithful transmission of genomic information requires tight spatiotemporal regulation of DNA replication factors. Posttranslational modifications, such as ubiquitylation, constitute a fast and effective mechanism to control such complex protein function. The AAA-ATPase CDC-48 plays an essential role in selective protein degradation triggered by ubiquitylation. While initial studies reported a crucial function of CDC-48 in the regulation of mitotic events, an essential role of the CDC-48/UFD-1/NPL-4 complex in DNA replication has been revealed in Caenorhabditis elegans (C. elegans) recently. Since the mechanistic details of CDC-48 activity remained to be elucidated, the identification of key substrates playing a vital role during DNA duplication is of major interest. This work describes a regulatory function of CDC-48 in the coordination of licensing and elongation events of DNA replication in C. elegans. In the licensing step of DNA replication, CDT-1 is loaded onto chromatin to subsequently promote the recruitment of relevant replication factors, including CDC-45 and the GINS complex. Throughout the elongation step CDC-45 and the GINS complex move with the replication fork, however, it is largely unknown how their chromatin association is controlled. CDC-48/UFD-1/NPL-4 deficient embryos stabilize the licensing factor CDT-1 exclusively on mitotic chromatin. Furthermore, worm embryos lacking cdc-48, ufd-1, or npl-4, show persistent chromatin association of CDC-45 and the GINS complex. Notably, the protein levels of CDC-45 and the GINS subunits SLD-5 and PSF-3 are not affected by ufd-1 and npl-4 (RNAi), suggesting a non-proteolytic regulation. Down-regulation of CDT-1 suppresses the chromatin association of the GINS complex in embryos disrupted for a functional CDC-48/UFD-1/NPL-4 complex. Hence, CDC-48 is supposed to orchestrate both, CDT-1 degradation and chromatin dissociation of the CDC-45/GINS complex. In conclusion, this work describes a novel role of the ubiquitin-selective chaperone CDC-48/UFD-1/NPL-4 in the context of chromatin associated processes. Elucidating the key substrates of CDC-48 during DNA replication illustrates a critical function in safeguarding genomic stability by an unexpected principle of target protein regulation

Clostridium difficile colitis in patients after kidney and pancreas-kidney transplantation

Limited data exist about Clostridium difficile colitis (CDC) in solid organ transplant patients. Between 1/1/99 and 12/31/02, 600 kidney and 102 pancreas–kidney allograft recipients were transplanted. Thirty-nine (5.5%) of these patients had CDC on the basis of clinical and laboratory findings. Of these 39 patients, 35 have information available for review. CDC developed at a median of 30 days after transplantation, and the patients undergoing pancreas–kidney transplantation had a slightly higher incidence of CDC than recipients of kidney alone (7.8% vs. 4.5%, P> 0.05). All but one patient presented with diarrhea. Twenty-four patients (64.9%) were diagnosed in the hospital, and CDC occurred during first hospitalization in 14 patients (40%). Treatment was with oral metronidazole (M) in 33 patients (94%)and M + oral vancomycin (M + V) in 2 patients. Eight patients had recurrent CDC, which occurred at a median of 30 days (range 15–314) after the first episode. Two patients (5.7%) developed fulminant CDC, presented with toxic megacolon, and underwent colectomy. One of them died; the other patient survived after colectomy. CDC should be considered as a diagnosis in transplant patients with history of diarrhea after antibiotic use, and should be treated aggressively before the infection becomes complicated

Complement deficiencies limit CD20 monoclonal antibody treatment efficacy in CLL

Monoclonal antibodies (MAbs) form a central part of chronic lymphocytic leukaemia (CLL) treatment. We therefore evaluated whether complement defects in CLL patients reduced the induction of complement-dependent cytotoxicity (CDC) by using anti-CD20 MAbs rituximab (RTX) and ofatumumab (OFA). Ofatumumab elicited higher CDC levels than RTX in all CLL samples examined, particularly in poor prognosis cohorts (11q− and 17p−). Serum sample analyses revealed that 38.1% of patients were deficient in one or more complement components, correlating with reduced CDC responses. Although a proportion of patients with deficient complement levels initially induced high levels of CDC, on secondary challenge CDC activity in sera was significantly reduced, compared with that in normal human serum (NHS; P<0.01; n=52). In addition, a high CLL cell number contributed to rapid complement exhaustion. Supplementing CLL serum with NHS or individual complement components, particularly C2, restored CDC on secondary challenge to NHS levels (P<0.0001; n=9). In vivo studies revealed that complement components were exhausted in CLL patient sera post RTX treatment, correlating with an inability to elicit CDC. Supplementing MAb treatment with fresh-frozen plasma may therefore maintain CDC levels in CLL patients with a complement deficiency or high white blood cell count. This study has important implications for CLL patients receiving anti-CD20 MAb therapy

Reasoning about Cardinal Directions between Extended Objects

Direction relations between extended spatial objects are important commonsense knowledge. Recently, Goyal and Egenhofer proposed a formal model, known as Cardinal Direction Calculus (CDC), for representing direction relations between connected plane regions. CDC is perhaps the most expressive qualitative calculus for directional information, and has attracted increasing interest from areas such as artificial intelligence, geographical information science, and image retrieval. Given a network of CDC constraints, the consistency problem is deciding if the network is realizable by connected regions in the real plane. This paper provides a cubic algorithm for checking consistency of basic CDC constraint networks, and proves that reasoning with CDC is in general an NP-Complete problem. For a consistent network of basic CDC constraints, our algorithm also returns a 'canonical' solution in cubic time. This cubic algorithm is also adapted to cope with cardinal directions between possibly disconnected regions, in which case currently the best algorithm is of time complexity O(n^5)

The dynamics of replication licensing in live Caenorhabditis elegans embryos

Accurate DNA replication requires proper regulation of replication licensing, which entails loading MCM-2-7 onto replication origins. In this paper, we provide the first comprehensive view of replication licensing in vivo, using video microscopy of Caenorhabditis elegans embryos. As expected, MCM-2-7 loading in late M phase depended on the prereplicative complex (pre-RC) proteins: origin recognition complex (ORC), CDC-6, and CDT-1. However, many features we observed have not been described before: GFP-ORC-1 bound chromatin independently of ORC-2-5, and CDC-6 bound chromatin independently of ORC, whereas CDT-1 and MCM-2-7 DNA binding was interdependent. MCM-3 chromatin loading was irreversible, but CDC-6 and ORC turned over rapidly, consistent with ORC/CDC-6 loading multiple MCM-2-7 complexes. MCM-2-7 chromatin loading further reduced ORC and CDC-6 DNA binding. This dynamic behavior creates a feedback loop allowing ORC/CDC-6 to repeatedly load MCM-2-7 and distribute licensed origins along chromosomal DNA. During S phase, ORC and CDC-6 were excluded from nuclei, and DNA was overreplicated in export-defective cells. Thus, nucleocytoplasmic compartmentalization of licensing factors ensures that DNA replication occurs only once

Fate specification and tissue-specific cell cycle control of the <i>Caenorhabditis elegans</i> intestine

Coordination between cell fate specification and cell cycle control in multicellular organisms is essential to regulate cell numbers in tissues and organs during development, and its failure may lead to oncogenesis. In mammalian cells, as part of a general cell cycle checkpoint mechanism, the F-box protein β-transducin repeat-containing protein (β-TrCP) and the Skp1/Cul1/F-box complex control the periodic cell cycle fluctuations in abundance of the CDC25A and B phosphatases. Here, we find that the Caenorhabditis elegans β-TrCP orthologue LIN-23 regulates a progressive decline of CDC-25.1 abundance over several embryonic cell cycles and specifies cell number of one tissue, the embryonic intestine. The negative regulation of CDC-25.1 abundance by LIN-23 may be developmentally controlled because CDC-25.1 accumulates over time within the developing germline, where LIN-23 is also present. Concurrent with the destabilization of CDC-25.1, LIN-23 displays a spatially dynamic behavior in the embryo, periodically entering a nuclear compartment where CDC-25.1 is abundant

Implementation experiences of NASTRAN on CDC CYBER 74 SCOPE 3.4 operating system

The implementation of the NASTRAN system on the CDC CYBER 74 SCOPE 3.4 Operating System is described. The flexibility of the NASTRAN system made it possible to accomplish the change with no major problems. Various sizes of benchmark and test problems, ranging from two hours to less than one minute CP time were run on the CDC CYBER SCOPE 3.3, Univac EXEC-8, and CDC CYBER SCOPE 3.4. The NASTRAN installation deck is provided

Computer program to determine pressure distributions and forces on blunt bodies of revolution

Program was written to include integration of surface pressure in order to obtain axial-force, normal-force, and pitching-moment coefficients. Program was written in CDC FORTRAN for the CDC-6600 computer system