Requirement of yeast fimbrin for actin organization and morphogenesis in vivo

The SAC6 gene was found by suppression of a yeast actin mutation. Its protein product, Sac6p (previously referred to as ABP67), was independently isolated by actin-filament affinity chromatography and colocalizes with actin in vivo. Thus Sac6p binds to actin in vitro, and functionally associates with actin structures involved in the development and maintenance of cell polarity in vivo. We report here that Sac6p is an actin-filament bundling protein 43% identical in amino-acid sequence to the vertebrate bundling protein fimbrin. This yeast fimbrin homologue contains two putative actin-binding regions homologous to domains of dystrophin, beta-spectrin, filamin, actin-gelation protein and alpha-actinin. Mutants lacking Sac6p do not form normal actin structures and are defective in morphogenesis. These findings demonstrate an in vivo role for the well-documented biochemical interaction between fimbrin and actin.     

Identifying and incorporating change in information systems

The difficulty of identifying future requirements and the inflexibility of information systems make what is normally called the maintenance process difficult and costly, and may lead to information systems failure or obsolescence. This paper addresses the problem in a number of ways. It discusses three techniques which might be used to help identify future requirements. Two of these techniques are drawn from other disciplines; the third is not used widely in the information systems domain. All the techniques have broader applicability than maintenance alone, in that they are concerned with reducing uncertainty. The paper outlines some tenets of good applications software design, drawn from a number of sources, which will facilitate change by making the software design more flexible. Finally, these tenets are incorporated into information systems design through a proposed modification of the information systems life cycle framework. This framework—part of a “code of good practice” for information systems developers—could be incorporated into information systems development methodologies.     

Characteristics of Columbia spotted frog ( Rana luteiventris ) oviposition sites in northeastern Oregon, USA

Several western ranid frogs possess a unique strategy of breeding communally over a short temporal window and reusing oviposition sites between years. However, little is published on the characteristics of oviposition sites selected by these explosive breeders. The Columbia spotted frog ( Rana luteiventris ) is native to northwestern North America and is of conservation concern in the southern portions of its range. As part of a study examining relationships between livestock grazing and R. luteiventris habitat, we assessed characteristics of the species' oviposition sites in 25 fishless ponds in northeastern Oregon. Oviposition sites were generally in shallow water (<25 cm) close to shore and tended to be in the northeastern portion of ponds. Oviposition sites were found more frequently over heavily vegetated substrates and in areas of less substrate slope and shade than random points in littoral zones. We did not quantify temperature differences within ponds, but the patterns we documented are consistent with preferential use of warmer microhabitats for oviposition.     

Insertional mutagenesis identifies multiple networks of cooperating genes driving intestinal tumorigenesis

The evolution of colorectal cancer suggests the involvement of many genes. To identify new drivers of intestinal cancer, we performed insertional mutagenesis using the Sleeping Beauty transposon system in mice carrying germline or somatic Apc mutations. By analyzing common insertion sites (CISs) isolated from 446 tumors, we identified many hundreds of candidate cancer drivers. Comparison to human data sets suggested that 234 CIS-targeted genes are also dysregulated in human colorectal cancers. In addition, we found 183 CIS-containing genes that are candidate Wnt targets and showed that 20 CISs-containing genes are newly discovered modifiers of canonical Wnt signaling. We also identified mutations associated with a subset of tumors containing an expanded number of Paneth cells, a hallmark of deregulated Wnt signaling, and genes associated with more severe dysplasia included those encoding members of the FGF signaling cascade. Some 70 genes had co-occurrence of CIS pairs, clustering into 38 sub-networks that may regulate tumor development.     

In vivo continuous electrochemical determination of dopamine release in rat neostriatum]

Polarographic micro-electrodes (carbon fiber, o. d. 8 micron) implanted in the Rat caudate nucleus, allowed a practically continuous in vivo monitoring (one measurement every 5 sec.) of the extra-cellular concentration of dopamine released by striatal dopaminergic terminals. Administration of amphetamine produced a reproducible increase of the oxidation current. This effect was suppressed after the selective degeneration of the striatal dopaminergic terminals following the injection of 6-OHDA into the substantia nigra or after inhibition of the synthesis of the amine by alpha methyl-p-tyrosin. After 5 hrs. this drug produced a 70% decrease of the oxidation current.     

Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I

Eukaryotic cells are dependent on their ability to translocate membraneous elements about the cytoplasm. In many cells long translocations of organelles are associated with microtubules. In other cases, such as the rapid cytoplasmic streaming in some algae, organelles appear to be propelled along actin filaments. It has been assumed, but not proven, that myosin produces these movements. We have tested vesicles from another eukaryotic cell for their ability to move on the exposed actin bundles of Nitella as an indiction that actin-based organelle movements may be a general property of cells. We found that organelles from Acanthamoeba castellanii can move along Nitella actin filaments. Here, we report two different experiments indicating that the single-headed non-polymerizable myosin isozyme myosin-I is responsible for this organelle motility. First, monoclonal antibodies to myosin-I inhibit movement, but antibodies that inhibit double-headed myosin-II do not. Second, approximately 20% of the myosin-I in homogenates co-migrates with motile vesicles during Percoll density-gradient ultracentrifugation. This is the first indication of a role for myosin-I within the cell and supports the suggestion of Albanesi et al. that myosin-I moves vesicles in this way.