The Fission Yeast S-Phase Cyclin Cig2 Can Drive Mitosis [preprint]

Commitment to mitosis is regulated by cyclin-dependent kinase (CDK) activity. In the fission yeast Schizosaccharomyces pombe, the major B-type cyclin, Cdc13, is necessary and sufficient to drive mitotic entry. Furthermore, Cdc13 is also sufficient to drive S phase, demonstrating that a single cyclin can regulate alternating rounds of replication and mitosis and providing the foundation of the quantitative model of CDK function. It has been assumed that Cig2, a B-type cyclin expressed only during S-phase and incapable of driving mitosis in wild-type cells, was specialized for S-phase regulation. Here, we show that Cig2 is capable of driving mitosis. Cig2/CDK activity drives mitotic catastrophe -- lethal mitosis in inviably small cells -- in cells that lack CDK inhibition by tyrosine-phosphorylation. Moreover, Cig2/CDK can drive mitosis in the absence of Cdc13/CDK activity. These results demonstrate that in fission yeast, not only can the presumptive M-phase cyclin drive S phase, but the presumptive S-phase cyclin can drive M phase, further supporting the quantitative model of CDK function. Furthermore, these results provide an explanation, previously proposed on the basis of computation analyses, for the surprising observation that cells expressing a single-chain Cdc13-Cdc2 CDK do not require Y15 phosphorylation for viability. Their viability is due to the fact that in such cells, which lack Cig2/CDK complexes, Cdc13/CDK activity is unable to drive mitotic catastrophe

The NASA MLAS Flight Demonstration - A Review of a Highly Successful Test

NASA has tested the Max Launch Abort System (MLAS) as a risk-mitigation design should problems arise with the baseline Orion spacecraft launch abort design. The Max in MLAS is not Maximum, but rather dedicated to Max Faget, The renowned NASA Spacecraft designer. In the fall of 2009, the mission was flown, with great success, from the NASA Wallops Flight Facility. The MLAS flight test vehicle prototype consists of a boost ring, coast ring, and the MLAS fairing itself, which houses an Orion Command Module (CM) boilerplate. The objective of the MLAS flight test is to reorient the fairing with the CM, weighing approximately 29,000 lbs and traveling 290 fps, 180 degrees to an orientation suitable for the release of the CM during a pad abort and low altitude abort. Although multiple parachute deployments are used in the MLAS flight test vehicle to complete its objective, there are only two parachute types employed in the flight test. Five of the nine parachutes used for MLAS are 27.6 ft DO ribbon parachutes, and the remaining four are standard G-12 cargo parachutes. This paper presents an overview of the 27.6 ft DO ribbon parachute system employed on the MLAS flight test vehicle for coast ring separation, fairing reorientation, and as drogue parachutes for the CM after separation from the fairing. Discussion will include: the process used to select this design, previously proven as a spin/stall recovery parachute; descriptions of all components of the parachute system; the minor modifications necessary to adapt the parachute to the MLAS program; the techniques used to analyze the parachute for the multiple roles it performs; a discussion of the rigging techniques used to interface the parachute system to the vehicle; a brief description of how the evolution of the program affected parachute usage and analysis; and a summary of the results of the flight test, including video of the flight test and subsequent summary analysis. . A discussion of the flight test which was highly successful as well as the flight test observations will be a significant portion of the review

A Review of the MLAS Parachute Systems

The NASA Engineering and Safety Center (NESC) is developing the Max Launch Abort System (MLAS) as a risk-mitigation design should problems arise with the baseline Orion spacecraft launch abort design. The Max in MLAS is dedicated to Max Faget, the renowned NASA spacecraft designer. The MLAS flight test vehicle consists of boost skirt, coast skirt and the MLAS fairing which houses a full scale boilerplate Orion Crew Module (CM). The objective of the flight test is to prove that the CM can be released from the MLAS fairing during pad abort conditions without detrimental recontact between the CM and fairing, achieving performance similar to the Orion launch abort system. The boost and coast skirts provide the necessary thrust and stability to achieve the flight test conditions and are released prior to the test -- much like the Little Joe booster was used in the Apollo Launch Escape System tests. To achieve the test objective, two parachutes are deployed from the fairing to reorient the CM/fairing to a heatshield first orientation. The parachutes then provide the force necessary to reduce the total angle of attack and body angular rates required for safe release of the CM from the fairing. A secondary test objective after CM release from the fairing is to investigate the removal of the CM forward bay cover (FBC) with CM drogue parachutes for the purpose of attempting to synchronously deploying a set of CM main parachutes. Although multiple parachute deployments are used in the MLAS flight test vehicle to complete its objective, there are only two parachute types employed in the flight test. Five of the nine parachutes used for MLAS are 27.6 ft D(sub 0) ribbon parachutes, and the remaining four are standard G-12 cargo parachutes. This paper presents an overview of the 27.6 ft D(sub 0) ribbon parachute system employed on the MLAS flight test vehicle for coast skirt separation, fairing reorientation, and as drogue parachutes for the CM after separation from the fairing. Discussion will include: the process used to select this design, previously proven as a spin/stall recovery parachute; descriptions of all components of the parachute system; the minor modifications necessary to adapt the parachute to the MLAS program; the techniques used to analyze the parachute for the multiple roles it performs; a discussion of the rigging techniques used to interface the parachute system to the vehicle; and a brief description of how the evolution of the program affected parachute usage and analysis. An overview of the Objective system, rationale for the MLAS approach and the future of the program will also be presented. We hope to have flight test results to report at the time of the Conference Presentation

On the Mechanism of Gene Amplification Induced under Stress in Escherichia coli

Gene amplification is a collection of processes whereby a DNA segment is reiterated to multiple copies per genome. It is important in carcinogenesis and resistance to chemotherapeutic agents, and can underlie adaptive evolution via increased expression of an amplified gene, evolution of new gene functions, and genome evolution. Though first described in the model organism Escherichia coli in the early 1960s, only scant information on the mechanism(s) of amplification in this system has been obtained, and many models for mechanism(s) were possible. More recently, some gene amplifications in E. coli were shown to be stress-inducible and to confer a selective advantage to cells under stress (adaptive amplifications), potentially accelerating evolution specifically when cells are poorly adapted to their environment. We focus on stress-induced amplification in E. coli and report several findings that indicate a novel molecular mechanism, and we suggest that most amplifications might be stress-induced, not spontaneous. First, as often hypothesized, but not shown previously, certain proteins used for DNA double-strand-break repair and homologous recombination are required for amplification. Second, in contrast with previous models in which homologous recombination between repeated sequences caused duplications that lead to amplification, the amplified DNAs are present in situ as tandem, direct repeats of 7–32 kilobases bordered by only 4 to 15 base pairs of G-rich homology, indicating an initial non-homologous recombination event. Sequences at the rearrangement junctions suggest nonhomologous recombination mechanisms that occur via template switching during DNA replication, but unlike previously described template switching events, these must occur over long distances. Third, we provide evidence that 3′-single-strand DNA ends are intermediates in the process, supporting a template-switching mechanism. Fourth, we provide evidence that lagging-strand templates are involved. Finally, we propose a novel, long-distance template-switching model for the mechanism of adaptive amplification that suggests how stress induces the amplifications. We outline its possible applicability to amplification in humans and other organisms and circumstances

Dietary Restriction Induced Longevity is Mediated by Nuclear Receptor NHR-62 in Caenorhabditis elegans

Dietary restriction (DR) extends lifespan in a wide variety of species, yet the underlying mechanisms are not well understood. Here we show that the C. elegans HNF4a- related nuclear hormone receptor NHR-62 is required for metabolic and physiologic responses associated with DR-induced longevity. nhr-62 mediates the longevity of eat- 2 mutants, a genetic mimetic of dietary restriction, and blunts the longevity response of DR induced by bacterial food dilution at low nutrient levels. Metabolic changes associated with DR, including decreased Oil Red O staining, increased autophagy, and changes in fatty acid composition are partly reversed by mutation of nhr-62. Expression profiles reveal that several hundred genes induced by DR depend on the activity of NHR-62, including a putative lipase required for the DR response. This study provides critical evidence that nuclear hormone receptors regulate the DR response, suggesting hormonal and metabolic control of life span

Size-Dependent Expression of the Mitotic Activator Cdc25 as a Mechanism of Size Control in Fission Yeast [preprint]

Proper cell size is essential for cellular function (Hall et al., 2004). Nonetheless, despite more than 100 years of work on the subject, the mechanisms that maintain cell size homeostasis are largely mysterious (Marshall et al., 2012). Cells in growing populations maintain cell size within a narrow range by coordinating growth and division. Bacterial and eukaryotic cells both demonstrate homeostatic size control, which maintains population-level variation in cell size within a certain range, and returns the population average to that range if it is perturbed (Marshall et al., 2012; Turner et al., 2012; Amodeo and Skotheim, 2015). Recent work has proposed two different strategies for size control: budding yeast has been proposed to use an inhibitor-dilution strategy to regulate size at the G1/S transition (Schmoller et al., 2015), while bacteria appear to use an adder strategy, in which a fixed amount of growth each generation causes cell size to converge on a stable average, a mechanism also suggested for budding yeast (Campos et al., 2014; Jun and Taheri-Araghi, 2015; Taheri-Araghi et al., 2015; Tanouchi et al., 2015; Soifer et al., 2016). Here we present evidence that cell size in the fission yeast Schizosaccharomyces pombe is regulated by a third strategy: the size dependent expression of the mitotic activator Cdc25. The cdc25 transcript levels are regulated such that smaller cells express less Cdc25 and larger cells express more Cdc25, creating an increasing concentration of Cdc25 as cell grow and providing a mechanism for cell to trigger cell division when they reach a threshold concentration of Cdc25. Since regulation of mitotic entry by Cdc25 is well conserved, this mechanism may provide a wide spread solution to the problem of size control in eukaryotes

Large-scale mapping of bioactive peptides in structural and sequence space

Health-enhancing potential bioactive peptide (BP) has driven an interest in food proteins as well as in the development of predictive methods. Research in this area has been especially active to use them as components in functional foods. Apparently, BPs do not have a given biological function in the containing proteins and they do not evolve under independent evolutionary constraints. In this work we performed a large-scale mapping of BPs in sequence and structural space. Using well curated BP deposited in BIOPEP database, we searched for exact matches in non-redundant sequences databases. Proteins containing BPs, were used in fold-recognition methods to predict the corresponding folds and BPs occurrences were mapped. We found that fold distribution of BP occurrences possibly reflects sequence relative abundance in databases. However, we also found that proteins with 5 or more than 5 BP in their sequences correspond to well populated protein folds, called superfolds. Also, we found that in well populated superfamilies, BPs tend to adopt similar locations in the protein fold, suggesting the existence of hotspots. We think that our results could contribute to the development of new bioinformatics pipeline to improve BP detection.Fil: Nardo, Agustina Estefania. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; ArgentinaFil: Añon, Maria Cristina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; ArgentinaFil: Parisi, Gustavo Daniel. Universidad Nacional de Quilmes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

A Novel 3-Hydroxysteroid Dehydrogenase That Regulates Reproductive Development and Longevity

A multidisciplinary approach identifies novel biochemical activities involved in the synthesisof C. elegans bile acid-like steroids, which act as hormones that regulate sterol metabolism and longevity

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

The dinB Operon and Spontaneous Mutation in Escherichia coli

Apparently conflicting data regarding the role of SOS-inducible, error-prone DNA polymerase IV (DinB) in spontaneous mutation are resolved by the finding that mutation is reduced by a polar allele with which dinB and neighboring yafN are deleted but not by two nonpolar dinB alleles. We demonstrate the existence of a dinB operon that contains four genes, dinB-yafN-yafO-yafP. The results imply a role for yafN, yafO, and/or yafP in spontaneous mutation