Structural study of immunoaffinity-purified DNA polymerase α-DNA primase complex from calf thymus

The DNA polymerase α-DNA primase complex was purified over 17 000-fold to near homogeneity from calf thymus using an immunoaffinity column. Sodium dodecyl sulfate gel electrophoresis revealed three polypeptides with molecular weights of 140, 50 and 47 kDa, in a ratio of 1:2:0.25. The complex showed a sedimentation coefficient of 9.7 S, a Stokes radius of 56 Å and a native molecular weight of 250–260 kDa. Taken together, the data suggest that the calf thymus dNA polymerase α-DNA primase complex is essentially a heterotrimer of large (140 kDa) and small (50 kDa) subunits in a ratio of 1:2, with a globular conformation. Electron-microscopic studies of the complex revealed a spherical particle of 120 Å in diameter, in agreement with the physicochemical results. The binding of the complex to DNA was also demonstrated.     

Evidence of tool-use by chimpanzees (Pan troglodytes schweinfurthii) for digging out a bee-nest in the Kahuzi-biega national park,Zaïre

Two sticks were found near a broken bee-nest ofMeliplebeia tanganyikae aff.nigrita Alfken in the Mt. Kahuzi region of Zaïre, and were thought to have been used by a chimpanzee or perhaps several chimpanzees to dig out the subterranean nest. Honey, larvae, and most of the nest had been eaten by them. We did not find any evidence to indicate tool-use by chimpanzees in the Masisi or Itebero-Utu regions, although stingless bees were observed and honey was eaten by chimpanzees in both regions. The sticks resembled in length and diameter those known to be employed for digging termite-mounds in south-west Cameroon and Equatorial Guinea. The tool-behavior of the chimpanzees observed at Mt. Kahuzi may be similar to that of those in central Africa, rather than of those in east Africa where digging-tools have yet to be found. Another possibility is that the chimpanzees have developed the digging-tools independently, based on the need to take animal protein in the Mt. Kahuzi region, where termite-mounds are rarely observed. Instead of seeking termites, they may have a stronger motivation to seek bee larvae, especially the larvae of stingless bees beneath the ground, than to the chimpanzees inhabiting lower or drier forests.     

Expression of tandem gene fusions in transgenic tobacco plants.

We have studied the expression of four sets of tandem gene fusions in transgenic tobacco plants. This was to determine if the problem of between-transformant variability in expression of introduced genes could be overcome by using a linked reference gene as a co-ordinately expressed control. Tandem gene fusions containing identical 5' flanking regions (SSU301-ocs with either SSU301-cat or SSU301-SSU911) were not co-ordinately expressed in the transgenic tobacco plants whereas the tandem gene fusions containing similar but not identical 5' flanking regions (SSU301-ocs with SSU911-cat or SSU911-SSU301) were co-ordinately expressed. The lack of co-ordinate expression of some of the tandem gene fusions appears to be partially explained by absence of the corresponding genomic DNA segments in the transgenic plants.     

Acyl coenzyme A: Phospholipid acyltransferases in porcine platelets discriminate between ω-3 and ω-6 unsaturated fatty acids

The properties of porcine platelet acyltransferases which catalyze the incorporation of unsaturated fatty acids into the 2 positions of phospholipids were compared with those of porcine liver microsomes and rat liver microsomes. There were significant differences in the relative rates of incorporation of acyl groups into phospholipids as catalyzed by the membranes from different species and organs. The 1-acylglycerophosphate acyltransferase system showed relatively broad specificity for saturated and unsaturated fatty acids, with 14- to 20-carbon chains, while unsaturated acyl-CoAs with 18- and 20-carbon chains were generally good substrates in the acylations of 1-acylglycerophosphocholine and 1-acylglycerophosphoinositol. ω-3 and ω-6 unsaturated fatty acids were recognized differently by different acyltransferase systems in platelets. When activities for combinations of ω-3 and ω-6 unsaturated acyl-CoAs with the same number of carbons and with similar number of double bonds were compared, ω-6 fatty acids were relatively more preferred substrates than ω-3 fatty acids for the 1-acylglycerophosphoinositol acyltransferase system as compared with 1-acylglycerophosphocholine acyltransferase system.     

Molecular Shape and Packing in Crystals of Soybean β-Amylase

The structure of soybean β-amylase in trigonal (P3₂21) crystals was determined at 4.5 Å resolution by X-ray crystallographic techniques using the isomorphous replacement method. X-Ray diffraction data were collected by the screened precession method for the native enzyme and two heavy atom derivatives. The shape of the enzyme molecule and the locations of mercurial binding are presented. The molecule appeared to be composed of two domains: the larger domain contains one mercurial site on its surface and the smaller domain has another mercurial site, which seemed to be the so-called essential sulfhydryl group. A distinct cleft formed between the domains near the latter sulfhydryl group may be a substrate binding region.     

Sclerotic Rings in Mosasaurs (Squamata: Mosasauridae): Structures and Taxonomic Diversity

Mosasaurs (Squamata: Mosasauridae) were a highly diverse, globally distributed group of aquatic lizards in the Late Cretaceous (98–66 million years ago) that exhibited a high degree of adaptation to life in water. To date, despite their rich fossil record, the anatomy of complete mosasaur sclerotic rings, embedded in the sclera of the eyeball, has not been thoroughly investigated. We here describe and compare sclerotic rings of four mosasaur genera, Tylosaurus, Platecarpus, Clidastes, and Mosasaurus, for the first time. Two specimens of Tylosaurus and Platecarpus share an exact scleral ossicle arrangement, excepting the missing portion in the specimen of Platecarpus. Furthermore, the exact arrangement and the total count of 14 ossicles per ring are shared between Tylosaurus and numerous living terrestrial lizard taxa, pertaining to both Iguania and Scleroglossa. In contrast, two species of Mosasaurus share the identical count of 12 ossicles and the arrangement with each other, while no living lizard taxa share exactly the same arrangement. Such a mosaic distribution of these traits both among squamates globally and among obligatorily aquatic mosasaurs specifically suggests that neither the ossicle count nor their arrangement played major roles in the aquatic adaptation in mosasaur eyes. All the mosasaur sclerotic rings examined consistently exhibit aperture eccentricity and the scleral ossicles with gently convex outer side. Hitherto unknown to any squamate taxa, one specimen of Platecarpus unexpectedly shows a raised, concentric band of roughened surface on the inner surface of the sclerotic ring. It is possible that one or both of these latter features may have related to adaptation towards aquatic vision in mosasaurs, but further quantitative study of extant reptilian clades containing both terrestrial and aquatic taxa is critical and necessary in order to understand possible adaptive significances of such osteological features.     

Comparisons of Acids in Smoke of Lamina and Midrib of Flue-cured Tobacco Leaves

Acids of lamina and midrib cigarette smoke were converted into trimethylsilyl derivatives and they were analyzed with glass capillary column gas chromatography. Then compositional differences of acids between lamina and midrib cigarette smoke were discussed. The concentrations of organic acids were higher for lamina cigarette smoke than for midrib cigarette smoke. Large concentration difference were found in formic, acetic, propionic, lactic, glycolic, furoic, benzoic, phenylacetic, fumalic and m-hydroxybenzoic acid. Succinic and methylsuccinic acid were similar in lamina smoke and in midrib smoke.

A large amount of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one(amino-carbonyl reaction product) was identified for the first time in lamina smoke.     

Giant Cocoon Formation in the Silkworm,Bombyx mori L., Topically Treated with Methyleneclioxyphenyl Derivatives

To analyze the genetic system of paralytic shellfish poisoning (PSP) toxin production in the dinoflagellate Alexandrium catenella, we examined toxin compositions and mating type of Fl progenies from crosses between algal strains having different toxin compositions. In all strains used, the mole percentage of their toxin composition did not significantly change in any growth phase, although total toxin levels increased rapidly in the early to middle exponential growth phase and then decreased by 95% in the stationary phase. One parental strain produced gonyautoxin (GTX) 4, and C4, while the other produced neosaxitoxin (neoSTX) and saxitoxin (STX) during all growth phases. Fl progenies showed one parental toxin composition and segregated independently with the mating type. These data suggest that A. catenella is a toxin producer and that Mendelian inheritance of toxin profiles occurs in the heterothallic dinoflagellate A. catenella.