Mutational and kinetic analyses of the GTPase-activating protein (GAP)-p21 interaction: the C-terminal domain of GAP is not sufficient for full activity.

The GTPase-activating protein (GAP) stimulates the GTPase reaction of p21 by 5 orders of magnitude such that the kcat of the reaction is increased to 19 s-1. Mutations of residues in loop L1 (Gly-12 and Gly-13), in loop L2 (Thr-35 and Asp-38), and in loop L4 (Gln-61 and Glu-63) influence the reaction in different ways, but all of these mutant p21 proteins still form complexes with GAP. The C-terminal domain of the human GAP gene product, GAP334, which comprises residues 714 to 1047, is 20 times less active than full-length GAP on a molar basis and has a fourfold lower affinity. This finding indicates that the N terminus of GAP containing the SH2 domains modifies the interaction between the catalytic domain and p21.     

Anther and tissue culture-induced grain chalkiness and associated variants in rice

Rice, Oryza sativa, plants regenerated from anther culture with and without in vitro selection pressure were evaluated for chalky seed. Progeny evaluated included 21 spontaneously doubled haploids selfed 4 times, progeny from plants regenerated from S-aminoethylcysteine resistant callus selfed 4 times and backcrosses of both types to the parental type. All lines with in vitro histories had higher seed chalkiness than the controls both in the intensity of chalkiness and in the number of seeds expressing the character. The full range of intensity and amount of chalkiness was expressed in the progeny. The average intensity of anther/tissue culture-derived progeny was 4–5, based on a scale of 1 (translucent) to 10 (fully opaque), and the average amount of chalkiness within plants sampled was 50–75 percent. The chalky characteristic is transmitted from parent to offspring into a range of identifiable F₂ segregants. Disclaimer statement Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the USDA, and does not imply its approval to the exclusion of other products that may also be suitable.     

Karyotype and C-banding inTrigonella sectionFoenumgraecum (Fabaceae)

The six species of the sectionFoenum-graecum ofTrigonella have the same chromosome number, 2n = 16.T. gladiata andT. cariensis have fairly symmetrical karyotypes, while those ofT. foenum-graecum, T. berythea, T. macrorrhyncha andT. cassia are asymmetrical. C-bands are present in all six species but the number of bands and their positive vary considerably among the species. The karyotype evidence suggests that none of the available species of theFoenum-graecum section can be considered as the wild progenitor of fenugreek.     

Synthesis and maturation of cross-reactive glycoprotein in fibroblasts deficient in arylsulfatase a activity

The biosynthesis of arylsulfatase A was studied in cultured fibroblasts by pulse-chase labeling with [2-³H]mannose; the enzyme was isolated by immunoprecipitation and denaturing polyacrylamide gel electrophoresis. In normal fibroblasts, and in fibroblasts from a patient with multiple sulfatase deficiency, the enzyme was synthesized as a glycoprotein of apparent molecular weight of 59,000; half of it was processed over a period of 4 days to M_r= 57,000. The precursor chain of M_r= 59,000 was secreted in the presence of 10 mM NH₄Cl. An immunoprecipitable glycoprotein of normal size was synthesized by fibroblasts from two unrelated patients with metachromatic leukodystrophy, but this material disappeared within twenty hours. In fibroblasts from an individual with pseudodeficiency of arylsulfatase A, the immunoprecipitable precursor glycoprotein was smaller (M_r= 56,000). The synthesis of cross-reactive proteins with altered properties supports the concept of allelic mutations as the genetic basis of metachromatic leukodystrophy and of arylsulfatase A pseudodeficiency.     

Ancient dominance of the Quercus calliprinos - Pistacia palaestina association in mediterranean Israel

Abstract. The natural Mediterranean maquis and forest vegetation of Israel is commonly considered to be composed mainly of four, roughly equal components: Pinus halepensis, deciduous oak, evergreen oak, and Ceratonia - Pistacia communities. They represent the past climax and subclimax of this region. Evidence accumulated from pollen analysis and wood remnant research in geological and archaeological excavations, as well as from written historical sources, shows that this view is wrong: the ancient vegetation in this area was dominated by Quercus calliprinos.     

Retinoic acid modulates the pattern of cell division in embryos ofLymnaea stagnalis (Mollusca)

All-trans retinoic acid is well known as a modulator of positional specification in vertebrate development. A similar mechanism may operate in molluscan development. Molluscan development is characterized by an invariant pattern of cell divisions, which allows the study of individual cells in the developing organism. Low concentrations of exogenous retinoic acid applied during gastrulation affect the cell division pattern in the early larval stage of the molluscLymnaea stagnalis. A few cells from the apical plate, a larval organ consisting of seven large cleavage-arrested cells, were induced by retinoic acid to resume cell division. They typically formed an area of proliferating small cells that resembles the adjacent areas of precursor cells of adult ectoderm. The identification of individual cells that are transformed by retinoic acid may provide new insights into the mechanisms underlying positional specification within the embryo.     

Lentil domestication: On the quality of evidence and arguments

The current views on lentil domestication are based on biological attributes of the wild progenitorLens culinaris ssp. orientalis and on assumptions which have never been tested. Seed dormancy, a major factor in the adaptation of ssp.orientalis to its natural habitat, makes it inappropriate for cultivation, because poor germination causes seed yield following cultivation to be equal to the amount of sown seeds. Higher yield, resulting from the evolution of a non-dormant type can be obtained only after five or six cycles of unprofitable cultivation. It is doubtful that incipient farmers would have undertaken such an endeavor without preexisting knowledge that non-dormant types could eventually be obtained. Experiments involving the sowing of wild lentil would have been much more successful if the non-dormant types were present in appreciable quantities in the seed stock. Establishment of that type in the natural population would have required a period of seven to eight years with favorable growing conditions allowing the non-dormant type to become widespread in the population, followed by massive predation by man reducing the hazard of a population explosion. The close similarity between isozyme profiles of the cultivated lentil and its wild progenitor indicates that lentil cultivation was attempted with seeds derived from different populations and in different areas.     

Crystallization and preliminary X-ray analysis of nonglycosylated tissue inhibitor of metalloproteinases-1, N30QN78Q TIMP-1

A nonglycosylated (N₃₀QN₇₈Q) form of the human tissue inhibitor of metalloproteinases, TIMP-1, has been prepared and crystallized in a form suitable for X-ray diffraction analysis. Small single crystals have been grown using sodium tartrate as a precipitant. The crystals are in space group P2₁, with cell dimensions a = 35.28, b = 53.95, c = 48.56, and β = 96.0°. There is a single molecule of TIMP-1 in the asymmetric unit. The crystals diffract to at least 2.3 Å resolution. Complete data have been collected to 2.9 Å and a search for heavymetal derivatives is in progress. © 1993 Wiley-Liss, Inc.     

Activity of single-stranded DNA endonucleases in mung bean is associated with cell division

A single-strand-specific endonuclease from mung bean sprouts is widely usedin molecular biology. However, the biological role of this enzyme is unknown. We studied the spatial and temporal activity of single-stranded DNA endonucleases in mung bean seedling by following enzyme activity that linearizes supercoiled plasmid DNA, a characteristic of this type of enzyme. The formation of a linear molecule from supercoiled DNA was found to occur in two distinguishable steps. The first, which involves introducing a nick into the supercoiled DNA and relaxing it, is very rapid and complete within a few seconds. The second step of cleaving the opposite strand to generate a unit-length linear duplex DNA is a relatively slow process. Analysis of the DNA cleavage sites showed the nuclease preferentially cuts supercoiled DNA at an AT-rich region. Varying levels of nuclease activity could be detected in different tissues of the mung bean seedling. The highest activity was in the root tip and was correlated with histone H1 kinase activity. This implies a link between nuclease activity and cell division. Induction of cell division in mung bean hypocotyls with auxin promoted formation of root primordia and considerably increased the activity of single-stranded DNA endonucleases. The nuclease activity and histone H1 kinase activity were reduced in mung bean cuttings treated with hydroxyurea, but not in cuttings treated with oryzalin. The potential function of single-stranded DNA endonucleases is discussed.