Effects of basic proteins of low molecular weight on the phosphohydrolase and phosphotransferase activities of microsomal glucose-6-phosphatase in adult monkey hepatocytes (author's transl)

The effects of histone 2A and some polycations on microsomal carbamylphosphate:D-glucose phosphotransferase and glucose-6-phosphate phosphohydrolase activities (D-glucose-6-phosphate phosphohydrolase, EC 3.1.3.9), have been investigated. 1. Histone 2A and polycations activate the two enzymic activities. At a constant cation concentration, this activation increases with the number of cationic groups per molecule. 2. Activation by histone 2A is related to its fixation on microsomal membranes. This fixation varies with quantities of histones and pH. 3. The nature of the interactions between histones and microsomal membranes is shown to be electrostatic, probably between the cationic groups of histones and the anionic group of membranous lipids. 4. Kinetic analysis reveal that histone 2A increases the maximal reaction velocity but does not affect the apparent Michaelis constant values for the substrates. 5. The role played by the cationic groups of histone 2A on the microsomal glucose 6-phosphatase, is discussed.     

Promotion of Flowering by DNA Base Analogues and Changes in Acid Phosphatase and Peroxidase Isozyme Composition in Dark-Grown Arabidopsis thaliana

A DNA base analogue, 5-bromodeoxyuridine (BUdR), promoted floweringof Arabidopsis thaliana in short and long photoperiods and evenin total darkness. The promotive effect of BUdR was nullifiedby thymidine which had a weak inhibitory effect by itself. AnotherDNA base analogue, 5-fluorodeoxyuridine (FUdR), inhibited theflowering at a low concentration (10^–8 M), but markedlyenhanced the promotive effect of BUdR if they were present togetherin the culture medium. In the flower-promoting medium containing both BUdR and FUdR,the number of acid phosphatase isozymes decreased temporarily,followed by an increase to the control level with a prolongedculture period. The number of peroxidase isozymes was greaterin plants grown in the medium with BUdR or BUdR $ FUdR thanin those without them. (Received October 22, 1987; Accepted March 25, 1988)     

Early development of the laboratory-reared flounder,Pleuronichthys cornutus

Fertilized eggs ofPleuronichthys cornutus were obtained by both artificial fertilization and natural spawning of laboratory-reared fish. The present paper describes in detail the early development of the fish and the rearing methods employed to provide basic information for mass production of this species. Eggs and sperm for artificial fertilization were obtained from adult fish caught in the Ariake Sound, Kyushu in November and December of 1984. Their maturation was successfully induced by intermuscular injection of pituitary homogenate of the silver carp,Hypophthalmichthys molitrix. Fertilized eggs were also obtained in 1985 by natural spawning of a broodstock kept in a tank for a year. Hatched larvae were fed successively with rotifers,Artemia nauplii and the harpacticoid copepod,Tigriopus japonicus and reared for 80 days. Ten thousand young fish of about 33 mm TL were obtained in 1984 and 1985 with the survival rate of about 17%. Ten developmental stages were defined on the basis of the morphological characteristics: A) newly hatched to 4 day old larvae, 2.7 to 4.1 mm TL (2.6 to 3.9 mmNL), yolk sac present; B) 4 to 16 day old larvae, 3.8 to 5.9 mm (3.6 to 5.6 mm), yolk resorbed, actively feeding on rotifers; C) 15 to 30 day old larvae, 6.3 to 8.3 mm (6.0 to 7.9 mm), notochord straight, hypural fin ray visible; D) 24 to 40 day old larvae, 6.7 to 9.2 mm (6.4 to 8.8 mm), caudal notochord upturned (45°); E) 28 to 45 day old larvae, 7.9 to 10.8 mm (7.5 to 10.3 mm), caudal notochord upturned (45°–90°); F) 32 to 50 day old larvae, 10.8 to 15.7 mm (8.8 to 12.8 mm BL), eyes symmetrical; G) 35 to 66 day old larvae, 13.4 to 20.0 mm (10.9 to 16.3 mm), eyes asymmetrical, but left eye not visible from the right side; H) 40 to 75 day old larvae, 13.8 to 26.2 mm (11.3 to 21.4 mm), the upper edge of left eye visible over top of the head from the right side; I) 46 to 89 day old larvae, 20.1 to 27.4 mm (16.4 to 22.4mm), left eye on the edge of the head and pupil visible from the right side; and J) juveniles of 51 day old or over, 23.6 mm or more (19.3 mm or more), metamorphosis completed. One to three inflections were found for relative growth of total length, eye diameter, upper jaw length, preanal length, and distance between the base of the pectoral fin and the anus against the notochord length or body length. Two inflections were found for body length (or notochord length)-body weight relationship. Most inflections appeared at the stages of D, F and J, corresponding to the body length of 8, 9–12 and 18–22 mm respectively.     

Light Activation of NADP-Malate Dehydrogenase in Guard Cell Protoplasts from Vicia faba L

Light-induced swelling of guard cell protoplasts (GCP) from Vicia faba was accompanied by increases in content of K⁺ and malate. DCMU inhibited the increase of K⁺ and malate, and consequently swelling.

Effect of light on the activity of selected enzymes that take part in malate formation was studied. When isolated GCP were illuminated, NADP-malate dehydrogenase (NADP-MDH) was activated, and the activity reached a maximum within 5 minutes. The enzyme activity underwent 5- to 6-fold increase in the light. Upon turning off the light, the enzyme was inactivated in 5 minutes NAD-MDH and phosphoenolpyruvate carboxylase (PEPC) were not influenced by light. The rapid light activation of NADP-MDH was inhibited by DCMU, suggesting that the enzyme was activated by reductants from the linear electron transport in chloroplasts. An enzyme localization study by differential centrifugation indicates that NADP-MDH is located in the chloroplasts, NAD-MDH in the cytosol and mitochondria, and PEPC in the cytosol. After light activation, the activity of NADP-MDH in guard cells was 10 times that in mesophyll cells on a chlorophyll basis. The physiological significance of light-dependent activation of NADP-MDH in guard cells is discussed in relation to stomatal movement.

     

Characteristics of structural proteins of infectious flacherie virus from the silkworm, Bombyx mori

Structural proteins and the characteristics of infectious flacherie virus (IFV) purified from the silkworm, Bombyx mori, are described. The purified IFV had four major structural proteins, which were detected only in high concentration gels of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and a few minor ones. Molecular weights of the major proteins were 35,200 (VP 1), 33,000 (VP 2), 31,200 (VP 3), and 11,600 (VP 4), and numbers per virion were 62, 57, 54, and 31, respectively. Amino acid compositions of VP 1, VP 2, and VP 3 were similar to each other but that of VP 4 was somewhat different. By isoelectric focusing and two-dimensional electrophoresis, high resolution of the structural proteins was obtained with silver staining. The isoelectric points of the four major proteins were determined as 7.7(VP 1), 6.7(VP 2), 4.8(VP 3), and 5.5(VP 4). This work is the first report on insect picornaviruses that presents some discriminative properties of each viral protein that was compared to those of mammalian picornaviruses.     

Transductional Analysis of the leu-thr Region of Chromosome of Serratia marcescens

Transductional analysis of the chromosomal linkage map of Serratia marcescens near leu revealed the following order of loci: ser4-thr3-pyr1-pdx2-leu1-azi8.     

Possible Involvement of Abscisic Acid in Increases in Activities of Two Vacuolar H+-Pumps in Barley Roots under Aluminum Stress

Levels of abscisic acid (ABA) in barley roots increased upontreatment with AlCl₃. Treatment with AlCl₃ or ABA increasedboth ATP-dependent and PP_i-dependent H⁺-pumping activities intonoplast-enriched membrane vesicles. Increase in the H⁺-pumpingactivities caused by aluminum stress could result from increasedlevels of ABA. ¹Present address: Department of Botany, Faculty of Science,Hirosaki University, Hirosaki, Aomori, 036 Japan     

Production and characterization of ligninolytic enzymes ofBjerkandera adusta grown on wood meal/wheat bran culture and production of these enzymes using a rotary-solid fermenter

Manganese peroxidase (MnP) and lignin peroxidase (LiP) were produced by growing a white-rot fungusBjerkandera adusta statically, on a wood meal/wheat bran culture in flasks. MnP and LiP reached their maximum activity after 6 and 19 days of inoculation, respectively. Both MnP and LiP are thought to be important enzymes in lignin biodegradation byB. adusta. Ion exchange chromatography showed thatB. adusta produced a single LiP and a single MnP enzyme in wood meal/wheat bran culture. These enzymes were separated and characterized. The molecular weight of MnP was 46,500 with a pl of 3.9. The molecular weight of LiP was estimated to be 47,000 with a pl of 3.5. Spectral analysis demonstrated that both enzymes are heme proteins. Production of these enzymes was also achieved using a rotarysolid culture fermenter. MnP, LiP and veratryl alcohol oxidase were produced byB. adusta in the fermenter.     

An immunological approach to quantitate RNA polymerases in plant cell extracts

A polyclonal antiserum was raised against highly purified RNA polymerase II from soybean embryos. Pure RNA polymerase II was fractionated on sodium dodecyl sulfate-polyacrylamide gels and transferred onto nitrocellulose sheets, incubated with the immune antiserum and then with iodinated protein A. Autoradiograms showed that the immune antiserum recognized all subunits of RNA polymerase II. Subunits 42, 27 and 16 kdalton were particularly reactive. Application of this transfer technique to protein extracts from soybean embryos or from cultured soybean cells allowed the identification of subunits of RNA polymerase II in the extracts. Analysis of the staining of the bands on the autoradiograms for increasing amounts of pure RNA polymerase II demonstrated that the transfer was quantitative, so that standard curves could be drawn to estimate the unknown amounts of enzyme in the extracts.Abbreviations DEAE diethylaminoethyl - SDS sodium dodecyl sulfate