Is functional manganese involved in hydrogen-peroxide-stimulated anomalous oxygen evolution in CACl2-washed photosystem II membranes?

When detergent-derived photosystem II (PSII) membranes are treated with CaCl₂ to remove the three extrinsic proteins associated with the O₂-evolving complex, the resulting membranes (CaPSII) can still catalyze water oxidation if sufficient Ca²⁺ and Cl^- are present. When CaPSII membranes are exposed to single turnover flashes on an O₂ rate electrode, anomalous O₂ is produced by the first two flashes. The addition of catalase to the membrane suspension completely inhibits O₂ produced by the first two flashes, but not by subsequent flashes. Exogenous H₂O₂ stimulates anomalous O₂ production by the first few flashes in CaPSII membranes, but not in control PSII membranes. Diuron (DCMU) does not inhibit H₂O₂-stimulated O₂ production by the first flash. However, it does inhibit the O₂ yield of all subsequent flashes, indicating that all flash-induced O₂ signals in CaPSII membranes are dependent on photosystem II electron transport. H₂O₂ stimulation of O₂ yields is inhibited in Tris-, heat-, and EDTA-(ethylenediaminetetraacetic acid)-treated CaPSII. In the presence of high salt, H₂O₂ (but not EDTA) treatment of CaPSII, extracts Mn functional in normal photosynthetic O₂ evolution. The addition of exogenous Mn²⁺ reconstitutes anomalous O₂ production in Tris-and H₂O₂/EDTA-treated CaPSII preparations but only in the presence of H₂O₂. Anomalous H₂O₂-stimulated O₂ production can be observed both with a Clark electrode (steady state) and an O₂ rate electrode (flash sequence). The mechanism involves electron donation from H₂O₂, mediated by free Mn²⁺, to PSII, and the 33-kDa extrinsic protein under some conditions can block this process. Since H₂O₂ can remove functional Mn from CaPSII membranes, its presence can convert functional Mn to the Mn²⁺ mediator state required for anomalous O₂ production. EDTA binds Mn in CaPSII disrupted by H₂O₂ and prevents anomalous O₂ evolution.Abbreviations CaPSII a PSII preparation washed with approximately 1M CaCl₂ - Chl chlorophyll - DCBQ 2,6-dichloro-p-benzoquinone - DCMU (diuron) 3-(3,4-dichlorophenyl)-1,1-dimethylurea - EDTA ethylenediaminetetraacetic acid - MES 2-[N-morpholino]-ethanesulfonic acid - PSII a detergent-derived photosystem II membrane preparation - RC reaction center - Tris tris(hydroxymethyl)-aminomethane - Y_n oxygen rate electrode flash yield resulting from the nth flash of a sequence of single turnover flashes of light Operated by the Midwest Research Institute for the U.S. Department of Energy under contract DE-AC02-83CH10093.     

Gene regulation during dedifferentiation in Dictyostelium discoideum

During development of Dictyostelium discoideum, cells acquire the capacity to rapidly recapitulate morphogenesis. Therefore, when cells at the loose aggregate stage are disaggregated and challenged to reaggregate, they do so in a tenth of the original time. If loose aggregate cells are disaggregated and resuspended in buffered dextrose solution (erasure medium), they retain the capacity of rapid recapitulation for 80 min, then completely lose this capacity in a single, synchronous step referred to as the "erasure event." The erasure event sets in motion a program of dedifferentiation during which cells lose developmentally acquired characteristics at different times. The erasure event is inhibited by the addition of 10(-4) M cAMP to erasure medium. The synthesis of 33 growth-associated polypeptides, the synthesis of 53 development-associated polypeptides, and the level of 2 development-associated RNAs have been monitored during the erasure program and in cultures inhibited from erasing by the addition of 10(-4) M cAMP. Growth-associated polypeptides begin to be resynthesized and development-associated polypeptides exhibit dramatic decreases in rate of synthesis at different times throughout the first 240 min in erasure medium. Inhibiting the erasure event with cAMP has no major effect in the resynthesis of the majority of growth-associated polypeptides. Only one growth-associated polypeptide, V28, is completely inhibited by cAMP, suggesting that it may play a role in the erasure process. In contrast, inhibiting the erasure event with cAMP has a marked effect on the synthesis of development-associated polypeptides, causing a dramatic reduction in the rate at which synthesis decreases for 6 polypeptides, and completely inhibits the decrease in the synthetic rate of 8 polypeptides. The two development-associated RNAs, 16G1 and 10C3, exhibit two distinctly different patterns of loss during erasure, but in both cases cAMP added at time zero of the erasure process dramatically retards or inhibits loss. In addition, when cAMP is added just prior to the erasure event, it inhibits the erasure event and stimulates a rapid increase in the level of 16G1 RNA back to the developmental level. The level of 16G1 RNA then remains at this level for at least 400 min. When cAMP is added after the erasure event, it causes a low, transient increase in the level of 16G1 RNA. These results are considered both in relation to the program of erasure, and in relation to the role of cAMP in the expression of developmental genes during the forward program of development.     

Circulating steroid hormones during rapid aggressive responses of territorial male mountain spiny lizards, Sceloporus jarrovi

Levels of aggression and circulating steroid hormones were monitored simultaneously in free-living male lizards following a staged territorial aggressive encounter with another male. In the first 15 min following the aggressive encounter, the frequency of territorial patroling and the frequency of agonistic and advertisement displays increased four- to fivefold in resident males. In most cases these increases persisted for at least 90 min after withdrawal of the intruder male and probably persisted for the entire day of the encounter. Blood samples collected at 15-min intervals revealed no changes in circulating levels of testosterone or corticosterone while this behavioral change was occurring. Thus, the increase in aggressive behavior that follows a male-male territorial encounter in this species does not appear to be mediated by simultaneous changes in circulating levels of these hormones. Interspecific comparisons suggest that interspecific variation in steroid hormone involvement in rapid aggressive responses may depend on the mating system and the extent of male parental care.     

Ortstreue und Genflu? bei Inselvogelarten: Eleonorenfalke (Falco eleonorae) und Gelbschnabelsturmtaucher (Calonectris diomedea)

Summary Young birds of both species return almost exclusively to their natal colony for breeding and breeders are higly faithful to both partner and territory in subsequent years. There is no or very limited emigration to other colonies which are 2–5 km distant. The consequences of the genetic isolation are discussed: In both species the tendency to form subspecies can be detected. High mortality rates are interpreted as a mean to eliminate any degenerated bird which could result from the close interbreeding in these small and isolated populations.     

Modulation of water and urea transport in human red cells: Effects of pH and phloretin

Summary It has previously been shown by Macey and Farmer (Biochim. Biophys. Acta 211:104–106, 1970) that phloretin inhibits urea transport across the human red cell membrane yet has no effect on water transport. Jennings and Solomon (J. Gen. Physiol. 67:381–397, 1976) have shown that there are separate lipid and protein binding sites for phloretin on the red cell membrane. We have now found that urea transport is inhibited by phloretin binding to the lipids with aK ₁ of 25±8 m in reason-able agreement with theK _D of 54±5 m for lipid binding. These experiments show that lipid/protein interactions can alter the conformational state of the urea transport protein. Phloretin binding to the protein site also modulates red cell urea transport, but the modulation is opposed by the specific stilbene anion transport inhibitor, DIDS (4,4-diisothiocyano-2,2-stilbene disulfonate), suggesting a linkage between the urea transport protein and band 3. Neither the lipid nor the protein phloretin binding site has any significant effect on water transport. Water transport is, however, inhibited by up to 30% in a pH-dependent manner by DIDS binding, which suggests that the DIDS/band 3 complex can modulate water transport.     

Distribution of calcium ATPase in the sarcoplasmic reticulum of fast- and slow-twitch muscles determined with monoclonal antibodies

Summary Four monoclonal antibodies against the calcium ATPase in sarcoplasmic reticulum (SR) of rabbit fast-twitch skeletal muscle were characterized using SDS-PAGE, Western blots and immunofluorescence. The ultrastructural distribution of the antigens was determined using post-embedding immunolabeling. The antibodies recognized the calcium ATPase in the SR but not in transverse (T-) tubule or plasma membranes. The antibody, D12, had the same binding affinity for the calcium ATPase from fast-twitch (rabbit sternomastoid) and slow-twitch (rabbit soleus) fibers and the affinity fell by 30% after fixation for electron microscopy in both types of muscle fiber. Ultrastructural studies revealed that the density of D12 antibody binding to the terminal cisternae membrane of extensor digitorum longus (edl) and sternomastoid fibers was on average seven times greater than in the slow-twitch soleus and semimembranosus fibers. Since the affinity of the ATPase for the antibody was the same in SR from fast- and slow-twitch muscles, the concentration of calcium ATPase in the terminal cisternae membrane of fast-twitch fibers was seven times greater than in slow-twitch fibers. This conclusion was supported by the fact that the concentration of calcium ATPase in light SR membranes was six times greater in SR from fast-twitch fibers than in SR from slow-twitch fibers. The results provide strong evidence that the different calcium accumulation rates in mammalian fast- and slow-twitch muscles are due to different concentrations of calcium ATPase molecules in the SR membrane.     

Recruitment of lysozyme as a major enzyme in the mouse gut: Duplication,divergence, and regulatory evolution

Summary Two major types of lysozymec (M and P) occur in the mouse genus,Mus, and have been purified from an inbred laboratory strain (C58/J) ofM. domesticus. They differ in physical, catalytic, and antigenic properties as well as by amino acid replacements at 6 of 49 positions in the amino-terminal sequence. Comparisons with four other mammalian lysozymesc of known sequence suggest that M and P are related by a gene duplication that took place before the divergence of the rat and mouse lineages. M lysozyme is present in most tissues; achieves its highest concentration in the kidney, lung, and spleen; and corresponds to the lysozyme partially sequenced before from another strain ofM. domesticus. InM. domesticus and several related species, P lysozyme was detected chiefly in the small intestine, where it is probably produced mainly by Paneth cells. A survey of M and P levels in 22 species of muroid rodents (fromMus and six other genera) of known phylogenetic relationships suggests that a mutation that derepressed the P enzyme arose about 4 million years ago in the ancestor of the housemouse group of species. Additional regulatory shifts affecting M and P levels have taken place along lineages leading to other muroid species. Our survey of 187 individuals of wild house mice and their closest allies reveals a correlation between latitude of origin and level of intestinal lysozyme.     

Multiple forms of male-specific simple repetitive sequences in the genusMus

Summary Previous reports indicate that in laboratory strains of mice, males are distinct from females in possession of repetitive DNA, notably devoid of Eco RI and Hae III sites and rich in the simple tetranucleotides GATA/GACA. We report here that such sequences originated in an ancestor common to laboratory mice,Mus hortulanus, M. spretus, and possibly alsoM. cookii. Interestingly, other male-specific satellite sequences were detected inM. caroli, M. cookii, M. saxicola, andM. minutoides. This novel satellite is also likely to be composed of simple repetitious sequences, but does not contain GATA and GACA. Thus, the Y chromosome appears to contain a disproportionately large amount of simple repetitious DNA. An attractive explanation for these results is that long tandem arrays of simple repeated sequences are generated at high frequency throughout the genome and that they are retained for a longer time on the Y chromosome due to the absence of homologous pairing at meiosis.     

Sterol Composition and Ecdysteroid Content of Eggs of the Root-knot Nematodes Meloidogyne incognita and M. arenaria

Free and esterified sterols of eggs of the root-knot nematodes Meloidogyne incognita races 2 and 3 and M. arenaria race 1 were isolated and identified by gas-liquid chromatography-mass spectrometry. The major sterols of eggs of each race were 24-ethylcholesterol (33.4-38.8% of total sterol), 24-ethylcholestanol (18.3-25.3%), 24-methylcholesterol (8.6-11.7%), 24-methylcholestanol (7.7-12.5%), and cholesterol (4.6-11.6%). Consequently, the major metabolic transformation performed by Meloidogyne females or eggs upon host sterols appeared to be saturation of the sterol nucleus. The free and esterified sterols of the same race did not differ appreciably, except for a slight enrichment of the steryl esters in cholesterol. Although the sterol composition of Meloidogyne eggs differed from that of other life stages of other genera of plant-parasitic nematodes, the three Meloidogyne races could not be distinguished from each other by their egg sterols. Ecdysteroids, compounds with hormonal function in insects, were not detected by radioimmunoassay in the Meloidogyne eggs either as free ecdysteroids or as polar conjugates.     

Regulation of Dipeptidyl Aminopeptidase I and Angiotensin Converting Enzyme Activities in Cultured Murine Brain Cells by Cortisol and Thyroid Hormone

Cultures of dissociated brain cells from 15-day-old fetal mice were grown in the presence and absence of 20 or 50 nM triiodothyronine (T3), 30 or 300 nM cortisol, and 30 nM cortisol plus 50 nM T3 added to chemically defined media or in media supplemented with 15% serum from control and hypothyroid calves. The specific activities of five lysosomal enzymes--N-acetyl galactosaminidase, beta-glucuronidase, beta-galactosidase, cathepsin B, and dipeptidyl aminopeptidase I (DAP-I)--were higher in cells grown in calf serum than in cells grown in defined media. Of these enzymes, only DAP-I was elevated in activity when the cells were grown in hypothyroid calf serum instead of control calf serum. Elevation of DAP-I activity was reversed by addition of 20 nM T3 to hypothyroid calf serum. The enzymatic properties of DAP-I were similar whether the cells were grown in control or hypothyroid calf serum and were similar to those reported for human fibroblasts and the purified enzyme. When the cells were grown in defined media, cortisol decreased the activities of all lysosomal enzymes, with 300 nM cortisol being more effective than 30 nM cortisol. Addition of 50 nM T3 to 30 nM cortisol decreased DAP-I activity more than 30 nM cortisol alone, but 50 nM T3 alone in defined media did not alter DAP-I levels. The reduction of DAP-I activity in these cells by T3 required cortisol, unidentified components in serum, or both.(ABSTRACT TRUNCATED AT 250 WORDS)