Site-directed mutational analysis for the ATP binding of DnaA protein. Functions of two conserved amino acids (Lys-178 and Asp-235) located in the ATP-binding domain of DnaA protein in vitro and in vivo

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, is activated by binding to ATP in vitro. We introduced site-directed mutations into two amino acids of the protein conserved among various ATP-binding proteins and examined functions of the mutated DnaA proteins, in vitro and in vivo. Both mutated DnaA proteins (Lys-178 --> Ile or Asp-235 --> Asn) lost the affinity for both ATP and ADP but did maintain binding activity for oriC. Specific activities in an oriC DNA replication system in vitro were less than one-tenth those of the wild-type protein. Assay of the generation of oriC sites sensitive to P1 nuclease, using the mutated DnaA proteins, revealed a defect in induction of the duplex opening at oriC. On the other hand, expression of each mutated DnaA protein in the temperature-sensitive dnaA46 mutant did not complement the temperature sensitivity. We suggest that Lys-178 and Asp-235 of DnaA protein are essential for the activity needed to initiate oriC DNA replication in vitro and in vivo and that ATP binding to DnaA protein is required for DNA replication-related functions.     

Studies on chitin. VI. Binding of metal cations

The adsorption abilities of chitin and its congeners with two series of degrees of deacetylation prepared by two different deacetylation procedures were compared. Among the polysaccharides obtained by the heterogeneous method, those with higher amino group content had higher adsorption ability. Plots of collection percentages versus amino group content, however, did not give straight lines. The plot for the chelation on the congeners that were prepared by homogeneous hydrolysis had a maximum at about 50% amino group content, and the value was higher than that for the sample with the highest amino group content, which was prepared by heterogeneous hydrolysis. These results suggest that the polysaccharides with about 50% amino group content obtained by the homogeneous procedure are potentially useful for the removal of metals.     

Heat shock-induced DNA relaxation in vitro by DNA gyrase of Escherichia coli in the presence of ATP

Genetic studies revealed that DNA gyrase seems to catalyze immediate and transient DNA relaxation after Escherichia coli cells are exposed to heat shock (Ogata, Y., Mizushima, T., Kataoka, K., Miki, T., and Sekimizu, K. (1994) Mol. Gen. Genet. 244, 451-455). We have now obtained biochemical evidence to support this hypothesis. DNA gyrase catalyzed an increase in the linking number of DNA and relaxation of negatively supercoiled DNA, under physiological concentrations of ATP. Analyses by gel filtration chromatography of each subunit revealed that DNA relaxation activity co-migrated with each subunit. The linking number of DNA increased as the temperature increased. Further, the reaction was inhibited by nalidixic acid or by oxolinic acid. Based on these results, we propose that DNA gyrase participates in a concerted reaction with DNA topoisomerases in the immediate relaxation of DNA in cells exposed to heat shock.     

Absorption of millimeter to submillimeter waves by atmospheric water molecules

Using the values of the rotational molecular parameters (including cencrifugal distortion terms) of the H₂ ¹⁶O molecule, which can explain 12 observed transitions below 800 GHz, all rotational energy levels with normalized Boltzmann factors larger than 5×10^?8 at 300°K are calculated. Probabilities of all possible electric dipole transitions among these states, 2277 lines, are calculated using the eigenfunctions thus obtained, and the permanent electric dipole moment of 1.8546 Debye. Assuming the single and full Lorentzian line forms, we calculated the absorption coefficient for millimeter to submillimeter region. Our result, using the single term Lorentzian line form, agrees quite well with experiment for 1 Torr of water vapor in 760 Torr air at 300°K.     

EFFECT OF BAFFLE ON TRAY UNDER OSCILLATING CONDITION

Effect of baffle on a tray under oscillating conditions has been examined experimentally. To reduce the degree of liquid mixing and liquid channeling under oscillating conditions, it is necessary to install the baffles on tray.

Experiments were conducted with the air-water system, ft was found that the pressure drop change can be expressed as a sinusoidal function and the period coincides with tray oscillation. The degree of liquid mixing decreases when the baffles are installed on tray and it also decreases with the increase of liquid flow rate. The pressure drop increases with the installation of baffle on tray.

The suitable baffle number on tray is from 4 to 8 and in this region, the degree of liquid mixing reaches minimum and the liquid holdup takes a maximum value. @KEYWORDS: Tray, Oscillating, Liquid mixing, Baffle, Pressure drop, Liquid holdup.     

Emissivity of chlorinated polyethylene

The emissivity of chlorinated polyethylene containing varying amounts of combined chlorine has been investigated. It was found that the emissivity of these compounds increases significantly with increase in chlorination.     

Aging and thermal stability of Mg/SiC and Mg/Y2O3 reflection multilayers in the 25-35 nm region

Reflection measurements in the 25-35 nm region were made for Mg/SiC and Mg/Y2O3 multilayers kept in a low-humidity atmosphere for 4 or 5 years. Aged Mg/SiC multilayers keep their reflectances, and the reflectance value at 31.2 nm is 0.44 at 10 degrees of the normal angle of incidence. Aged Mg/Y2O3 multilayers change reflectance as top layer materials, and the best value at 30.1 nm is 0.40 at 10 degrees. Reflection measurements are also made for Mg-based multilayers that are annealed from room temperature to 400 degrees C at 50 degrees C intervals. Both multilayers keep their reflectance at annealing temperatures of 200 degrees C. These results suggest that both Mg-based multilayers can be applied to practical optics.