Expression in mammalian cells of a cloned gene encoding murine DNA methyltransferase

Mammalian DNA cytosine-5-methyltransferase (MTase, EC 2.1.1.37) is an essential component for establishing and maintaining cell-type specific methylation patterns in the genome. The cDNAfor the murine enzyme was previously cloned in segments. We have reconstructed the entire gene, encoding a protein of 1517 amino acids, from a set of overlapping CDNA clones. We report the assembly of two expression constructs in bacterial/mammalian shuttle vectors. Transcription in the first construct (pEMT) is driven by the cytomegalovirus enhancer/promoter and encodes a fusion protein with 15 additional aa at the N terminus, while the second construct (pJMT) is driven by the simian virus 40 early promoter/enhancer upstream from the natural ATG codon. Immunofluorescence microscopy and immunoblot analysis have shown that both constructs direct the synthesis of MTase in COS-1 cells. Enzyme activity in whole-cell lysates of transfected COS-1 cells transfected with pEMT and pJMT are on average tenfold and fivefold higher than in control, respectively. The specific activities of the recombinant and endogenous mouse-cell enzyme are similar. These expression constructs will be of use in studies of DNA methylation in mammals

Cuisine and Identity in Contemporary Japan

Anthropolog

DNA methylation and DNA methyltransferases

The prevailing views as to the form, function, and regulation of genomic methylation patterns have their origin many years in the past, at a time when the structure of the mammalian genome was only dimly perceived, when the number of protein-encoding mammalian genes was believed to be at least five times greater than the actual number, and when it was not understood that only ~10% of the genome is under selective pressure and likely to have biological function. We use more recent findings from genome biology and whole-genome methylation profiling to provide a reappraisal of the shape of genomic methylation patterns and the nature of the changes that they undergo during gametogenesis and early development. We observe that the sequences that undergo deep changes in methylation status during early development are largely sequences without regulatory function. We also discuss recent findings that begin to explain the remarkable fidelity of maintenance methylation. Rather than a general overview of DNA methylation in mammals (which has been the subject of many reviews), we present a new analysis of the distribution of methylated CpG dinucleotides across the multiple sequence compartments that make up the mammalian genome, and we offer an updated interpretation of the nature of the changes in methylation patterns that occur in germ cells and early embryos. We discuss the cues that might designate specific sequences for demethylation or de novo methylation during development, and we summarize recent findings on mechanisms that maintain methylation patterns in mammalian genomes. We also describe the several human disorders, each very different from the other, that are caused by mutations in DNA methyltransferase genes

Tradition and Japanese Social Organization: Institutional Development in a Tokyo Neighborhood

Anthropolog

Scalable System Scheduling for HPC and Big Data

In the rapidly expanding field of parallel processing, job schedulers are the "operating systems" of modern big data architectures and supercomputing systems. Job schedulers allocate computing resources and control the execution of processes on those resources. Historically, job schedulers were the domain of supercomputers, and job schedulers were designed to run massive, long-running computations over days and weeks. More recently, big data workloads have created a need for a new class of computations consisting of many short computations taking seconds or minutes that process enormous quantities of data. For both supercomputers and big data systems, the efficiency of the job scheduler represents a fundamental limit on the efficiency of the system. Detailed measurement and modeling of the performance of schedulers are critical for maximizing the performance of a large-scale computing system. This paper presents a detailed feature analysis of 15 supercomputing and big data schedulers. For big data workloads, the scheduler latency is the most important performance characteristic of the scheduler. A theoretical model of the latency of these schedulers is developed and used to design experiments targeted at measuring scheduler latency. Detailed benchmarking of four of the most popular schedulers (Slurm, Son of Grid Engine, Mesos, and Hadoop YARN) are conducted. The theoretical model is compared with data and demonstrates that scheduler performance can be characterized by two key parameters: the marginal latency of the scheduler $t_s$ and a nonlinear exponent $\alpha_s$. For all four schedulers, the utilization of the computing system decreases to < 10\% for computations lasting only a few seconds. Multilevel schedulers that transparently aggregate short computations can improve utilization for these short computations to > 90\% for all four of the schedulers that were tested.Comment: 34 pages, 7 figure

Performance Measurements of Supercomputing and Cloud Storage Solutions

Increasing amounts of data from varied sources, particularly in the fields of machine learning and graph analytics, are causing storage requirements to grow rapidly. A variety of technologies exist for storing and sharing these data, ranging from parallel file systems used by supercomputers to distributed block storage systems found in clouds. Relatively few comparative measurements exist to inform decisions about which storage systems are best suited for particular tasks. This work provides these measurements for two of the most popular storage technologies: Lustre and Amazon S3. Lustre is an open-source, high performance, parallel file system used by many of the largest supercomputers in the world. Amazon's Simple Storage Service, or S3, is part of the Amazon Web Services offering, and offers a scalable, distributed option to store and retrieve data from anywhere on the Internet. Parallel processing is essential for achieving high performance on modern storage systems. The performance tests used span the gamut of parallel I/O scenarios, ranging from single-client, single-node Amazon S3 and Lustre performance to a large-scale, multi-client test designed to demonstrate the capabilities of a modern storage appliance under heavy load. These results show that, when parallel I/O is used correctly (i.e., many simultaneous read or write processes), full network bandwidth performance is achievable and ranged from 10 gigabits/s over a 10 GigE S3 connection to 0.35 terabits/s using Lustre on a 1200 port 10 GigE switch. These results demonstrate that S3 is well-suited to sharing vast quantities of data over the Internet, while Lustre is well-suited to processing large quantities of data locally.Comment: 5 pages, 4 figures, to appear in IEEE HPEC 201

Abnormal X chromosome inactivation and sex-specific gene dysregulation after ablation of FBXL10

BACKGROUND: Almost all CpG-rich promoters in the mammalian genome are bound by the multidomain FBXL10 protein (also known as KDM2B, JHDM1B, CXXC2, and NDY1). FBXL10 is expressed as two isoforms: FBXL10-1, a longer form that contains an N-terminal histone demethylase domain with C-terminal F-box, CXXC, PHD, RING, and leucine-rich repeat domains, and FBXL10-2, a shorter form that initiates at an alternative internal exon and which lacks the histone demethylase domain but retains all other annotated domains. Selective deletion of Fbxl10-1 had been reported to produce a low penetrance and variable phenotype; most of the mutant animals were essentially normal. We constructed mutant mouse strains that were either null for Fbxl10-2 but wild type for Fbxl10-1 or null for both Fbxl10-1 and Fbxl10-2. RESULTS: Deletion of Fbxl10-2 (in a manner that does not perturb expression of Fbxl10-1) produced a phenotype very different from the Fbxl10-1 mutant, with craniofacial abnormalities, neural tube defects, and increased lethality, especially in females. Mutants that lacked both FBXL10-1 and FBXL10-2 showed embryonic lethality and even more extreme sexual dimorphism, with more severe gene dysregulation in mutant female embryos. X-linked genes were most severely dysregulated, and there was marked overexpression of Xist in mutant females although genes that encode factors that bind to Xist RNA were globally downregulated in mutant female as compared to male embryos. CONCLUSIONS: FBXL10 is the first factor shown to be required both for the normal expression and function of the Xist gene and for normal expression of proteins that associate with Xist RNA; it is proposed that FBXL10 coordinates the expression of Xist RNA with proteins that associate with this RNA. The function of FBXL10 is largely independent of the histone demethylase activity of the long form of the protein. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13072-016-0069-1) contains supplementary material, which is available to authorized users

Enabling On-Demand Database Computing with MIT SuperCloud Database Management System

The MIT SuperCloud database management system allows for rapid creation and flexible execution of a variety of the latest scientific databases, including Apache Accumulo and SciDB. It is designed to permit these databases to run on a High Performance Computing Cluster (HPCC) platform as seamlessly as any other HPCC job. It ensures the seamless migration of the databases to the resources assigned by the HPCC scheduler and centralized storage of the database files when not running. It also permits snapshotting of databases to allow researchers to experiment and push the limits of the technology without concerns for data or productivity loss if the database becomes unstable.Comment: 6 pages; accepted to IEEE High Performance Extreme Computing (HPEC) conference 2015. arXiv admin note: text overlap with arXiv:1406.492

Considerations for Soybean Insecticides and Fungicides

Several Iowa agribusinesses are offering soybean growers pest management plans that include applications of fungicide and insecticide. Although combining an insecticide and fungicide may be convenient, the results from our 2008 Pesticide Stewardship trials suggest this is a convenience that may not pay off