Human cytomegalovirus DNA: BamHI, EcoRI and PstI restriction endonuclease cleavage maps

The cloned HindIII fragments of human cytomegalovirus (HCMV) strain AD169 DNA were mapped with respect to the BamHI, EcoRI and PstI restriction endonuclease cleavage sites. Composite restriction endonuclease cleavage maps for the entire virus genome were constructed using the previously established linkages between the HindIII fragments.     

On the mechanism of photosynthetic electron transfer in Rhodopseudomonas capsulata and Rhodopseudomonas sphaeroides

(1) Current models for the mechanism of cyclic electron transport in Rhodopseudomonas sphaeroides and Rhodopseudomonas capsulata have been investigated by observing the kinetics of electron transport in the presence of inhibitors, or in photosynthetically incompetent mutant strains. (2) In addition to its well-characterized effect on the Rieske-type iron sulfur center, 5-(n-undecyl)-6-hydroxy-4,7-dioxobenzothiazole (UHDBT) inhibits both cytochrome b₅₀ and cytochrome b_?90 reduction induced by flash excitation in Rps. sphaeroides and Rps. capsulata. The concentration dependency of the inhibition in the presence of antimycin (approx. 2.7 mol UHDBT/mol reaction center for 50% inhibition of extent) is very similar to that of its inhibition of the antimycin-insensitive phase of ferricytochrome c re-reduction. UHDBT did not inhibit electron transfer between the reduced primary acceptor ubiquinone (Q^?_I) and the secondary acceptor ubiquinone (Q_II) of the reaction center acceptor complex. A mutant of Rps. capsulata, strain R126, lacked both the UHDBT and antimycin-sensitive phases of cytochrome c re-reduction, and ferricytochrome b₅₀ reduction on flash excitation. (3) In the presence of antimycin, the initial rate of cytochrome b₅₀ reduction increased about 10-fold as the E_h(7.0) was lowered below 180 mV. A plot of the rate at the fastest point in each trace against redox potential resembles the Nernst plot for a two-electron carrier with E_m(7.0) ≈ 125 ± 15 mV. Following flash excitation there was a lag of 100–500 μs before cytochrome b₅₀ reduction began. However, there was a considerably longer lag before significant reduction of cytochrome c by the antimycin-sensitive pathway occurred. (4) The herbicide ametryne inhibited electron transfer between Q^?_I and Q_II. It was an effective inhibitor of cytochrome b₅₀ photoreduction at E_h(7.0) 390 mV, but not at E_h(7.0) 100 mV. At the latter E_h, low concentrations of ametryne inhibited turnover after one flash in only half of the photochemical reaction centers. By analogy with the response to o-phenanthroline, it is suggested that ametryne is ineffective at inhibiting electron transfer from Q^?_I to the secondary acceptor ubiquinone when the latter is reduced to the semiquinone form before excitation. (5) At E_h(7.0) > 200 mV, antimycin had a marked effect on the cytochrome b₅₀ reduction-oxidation kinetics but not on the cytochrome c and reaction center changes or the slow phase III of the electrochromic carotenoid change on a 10-ms time scale. This observation appears to rule out a mechanism in which cytochrome b₅₀ oxidation is obligatorily and kinetically linked to the antimycin-sensitive phase of cytochrome c reduction in a reaction involving transmembrane charge transfer at high E_h values. However, at lower redox potentials, cytochrome b₅₀ oxidation is more rapid, and may be linked to the antimycin-sensitive reduction of cytochrome c. (6) It is concluded that neither a simple linear scheme nor a simple Q-cycle model can account adequately for all the observations. Future models will have to take account of a possible heterogeneity of redox chains resulting from the two-electron gate at the level of the secondary quinone, and of the involvement of cytochrome b_?90 in the rapid reactions of the cyclic electron transfer chain     

Toxoplasma gondii: Purine synthesis and salvage in mutant host cells and parasites

Toxoplasma gondii, growing exponentially in heavily infected mutant Chinese hamster ovary cells that had a defined defect in purine biosynthesis, did not incorporate [U-¹⁴C]glucose or [¹⁴C]formate into the guanine or adenine of nucleic acids. Intracellular parasites therefore must be incapable of synthesizing purines and depend on their host cells for them. Extracellular parasites, which are capable of limited DNA and RNA synthesis, efficiently incorporated adenosine nucleotides, adenosine, inosine, and hypoxanthine into their nucleic acids; adenosine 5′-monophosphate was the best utilized precursor. Extracellular parasites incubated with ATP labeled with ³H in the purine base and ³²P in the α-phosphate incorporated the purine ring 50-fold more efficiently than they did the α-phosphate. Thus, ATP is largely degraded to adenosine before it can be used by T. gondii for nucleic acid synthesis. Two pathways for the conversion of adenosine to nucleotides appear to exist, one involving adenosine kinase, the other hypoxanthine—guanine phosphoribosyl transferase. In adenosine kinase-less mutant parasites, the efficiency of incorporation of ATP or adenosine was reduced by 75%, which indicates the adenosine kinase pathway was predominant. Extracellular parasites incorporated ATP into both the adenine and the guanine of their nucleic acids, so ATP from the host cell could supply the entire purine requirement of T. gondii. However, ATP generated by oxidative phosphorylation in the host cell is not essential for parasites because they grew normally in a cell mutant that was deficient in aerobic respiration and almost completely dependent upon glycolysis.     

Mg2+ restores membrane potential in rat liver mitochondria deenergized by Ca2+ and phosphate movements

Cellular ornithine biosynthesis could be expected to play a significant role in putrescine formation and hence in growth. Two enzymes are involved in ornithine biosynthesis: arginase and transamidinase. These enzyme activities were studied in two human melanoma cell lines differing in their K_m of diamine oxidase for putrescine and in their tumorigenicity in nude mice. Arginase activity accounts for the majority of ornithine formed in the highly tumorigenic cell line, while the majority of ornithine is derived from transamidinase action in the poorly tumorigenic cell line, with concomitant formation of methyl guanidine, a potent inhibitor of diamine oxidase.     

Segmental flexibility of ribosomal protein S1 bound to ribosomes and Q beta-replicase

The protonization pattern of the endogenous donor component D₁ which feeds electrons directly into chl-a⁺_II has been analyzed in Tris-washed inside-out thylakoids with the aid of appropriate pH-indicators. It was found that under repetitive flash excitation the amount of protons released is proportional to the extent of D₁-oxidation, depending on the time between the flashes. The kinetics of D₁-oxidation (being practically the same as in normal Tris-washed chloroplasts) are faster than the proton release by two orders of magnitude. The results lead to the conclusion that D₁ is protonized in the reduced state with pK(D^ox₁) < 5 and becomes deprotonized in the oxidized state with pK(D^red₁) ? 8. The proton release is kinetically limited by a transport barrier. Implications on the interpretation of the proton release pattern in preparation with intact water oxidation are discussed.     

Kinetics of nitrogen and phosphorus release in varying food supplies byDaphnia magna

Rates of nitrogen and phosphorus release from individualDaphnia magna were determined by measuring ammonia and soluble reactive phosphorus in successive 10-min incubations in small (0.05 ml) vessels after the animals were removed from their food. Release rates of both nutrients were generally highest initially and decreased with time after removal. The ratio of nitrogen to phosphorus released increased with time after animals were removed from an artificial detritus/bacterial food; ratios were lower and changed with time less for animals fed algae. These data suggest errors may be introduced by assumptions of constant stoichiometry for nutrient release in varying environments.GLERL Contribution No. 268     

Deepwater sediments and trophic conditions in Florida lakes

Sediment cores were taken from near maximum depth in 15 Florida lakes representing a wide range of trophic conditions. Chemical analyses of surface sediments showed Al, Fe, and Ca to be the most abundant elements in all samples, and the ratio of Al to Ca to be smaller for eutrophic lakes. Sediment organic matter increased with trophic state, as did the degree to which it was enriched in nitrogen. Corresponding sediment C/N ratios decreased with increasing lake trophic state and showed significant negative correlation with chlorophylla, total N, and total P in the water column. Concentrations of sedimentary chlorophyll derivatives showed some relation to trophic state but differences in basin morphometry hinder its use as an inter-lake index of chlorophyll production.     

Excretion of total phosphorus,dissolved reactive phosphorus,ammonia, and urea by Lake MichiganMysis relicta

Rates of nutrient release byMysis relicta in Lake Michigan were measured on five nights at a 45-m station near Milwaukee, Wisconsin, U.S.A., in the summer of 1977. Nocturnal vertical migrations of the mysids were monitored with both echosounder tracings and vertical net tows. Estimates of the total areal dry mass of the mysids ranged from 600 to 1 820 mg m–2. Rates of release of dissolved reactive phosphorus, total phosphorus, ammonia, and urea were measured in dark incubations on shipboard. Excretion experiments were initiated immediately after mysids were collected from each of several vertical net hauls. The depths of maximum mysid densities corresponded approximately with a deep phytoplankton peak located in the vicinity of the thermocline. Semiquantitative demands for N and P by phytoplankton within this peak were obtained from¹⁴C estimates of primary production from a previous study, assuming a constant C:N:P ratio for the algae. These algal nutrient demands were compared to potential N and P release by the mysids to obtain a first approximation of the relative rates of nutrient supply and demand for the field phytoplankton populations. Our analysis indicates that mysids may directly supply about 1–10% of the daily N and P demands of the phytoplankton in the deep peak. However, indirect interactions betweenMysis relicta and other organisms, such as small zooplankton and fishes, could be major factors in nutrient recycling within the metalimnion and subthermocline region of Lake Michigan.Contribution No. 238, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Wisconsin.Contribution No. 238, Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Wisconsin.     

恐鱼类的新材料

本文记述了采自云南盘溪的一恐鱼类的新材料,代表该类的一新种。这一化石的发现和记述,对于地层的对比和了解该类化石的地理分布均有一定意义。