Dynamics of amino acid redistribution in the carnivorous Venus flytrap (Dionaea muscipula) after digestion of 13C/15N‐labelled prey

• Amino acids represent an important component in the diet of the Venus flytrap (Dionaea muscipula), and supply plants with much needed nitrogen resources upon capture of insect prey. Little is known about the significance of prey‐derived carbon backbones of amino acids for the success of Dionaea's carnivorous life‐style.
• The present study aimed at characterizing the metabolic fate of ¹⁵N and ¹³C in amino acids acquired from double‐labeled insect powder. We tracked changes in plant amino acid pools and their δ¹³C‐ and δ¹⁵N‐signatures over a period of five weeks after feeding, as affected by contrasting feeding intensity and tissue type (i.e., fed and non‐fed traps and attached petioles of Dionaea).
• Isotope signatures (i.e., δ¹³C and δ¹⁵N) of plant amino acid pools were strongly correlated, explaining 60% of observed variation. Residual variation was related to contrasting effects of tissue type, feeding intensity and elapsed time since feeding. Synthesis of nitrogen‐rich transport compounds (i.e., amides) during peak time of prey digestion increased ¹⁵N‐ relative to ¹³C‐ abundances in amino acid pools. After completion of prey digestion, ¹³C in amino acid pools was progressively exchanged for newly fixed ¹²C. The latter process was most evident for non‐fed traps and attached petioles of plants that had received ample insect powder.
• We argue that prey‐derived amino acids contribute to respiratory energy gain and loss of ¹³CO₂ during conversion into transport compounds (i.e., 2 days after feeding), and that amino‐nitrogen helps boost photosynthetic carbon gain later on (i.e., 5 weeks after feeding).

     

Reduced Activity of Geranylgeranyl Reductase Leads to Loss of Chlorophyll and Tocopherol and to Partially Geranylgeranylated Chlorophyll in Transgenic Tobacco Plants Expressing Antisense RNA for Geranylgeranyl Reductase

The enzyme geranylgeranyl reductase (CHL P) catalyzes the reduction of geranylgeranyl diphosphate to phytyl diphosphate. We identified a tobacco (Nicotiana tabacum) cDNA sequence encoding a 52-kD precursor protein homologous to the Arabidopsis and bacterial CHL P. The effects of deficient CHL P activity on chlorophyll (Chl) and tocopherol contents were studied in transgenic plants expressing antisense CHL P RNA. Transformants with gradually reduced Chl P expression showed a delayed growth rate and a pale or variegated phenotype. Transformants grown in high (500 μmol m⁻² s⁻¹; HL) and low (70 μmol photon m⁻² s⁻¹; LL) light displayed a similar degree of reduced tocopherol content during leaf development, although growth of wild-type plants in HL conditions led to up to a 2-fold increase in tocopherol content. The total Chl content was more rapidly reduced during HL than LL conditions. Up to 58% of the Chl content was esterified with geranylgeraniol instead of phytol under LL conditions. Our results indicate that CHL P provides phytol for both tocopherol and Chl synthesis. The transformants are a valuable model with which to investigate the adaptation of plants with modified tocopherol levels against deleterious environmental conditions.     

Enzymbestimmungen in Erythrocyten bei Kindern mit Down-Syndrom

Zusammenfassung Bei 20 Kleinkindern mit gesicherter freier Trisomie 21 wurden die Aktivitäten von 21 Enzymen des Energie- und des Glutathionstoffwechsels in isolierten Erythrocyten gemessen. Die Aktivitäten der Fructose-6-phosphatkinase, der Lactatdehydrgenase, der Glucose-6-phosphatdehydrogenase, der Glutathionreduktase (NADP) und der Glutamat-Oxalacetat-Transaminase wurden gegenüber gleichaltrigen Kontrollen signifikant erhöht gefunden. Lediglich die etwa 50%ige Steigerung der Fructose-6-phosphatkinase-Aktivität kann im Sinne eines Gen-Dosis-Effektes gedeutet werden.

---

Enzyme activities in the erythrocytes of children with Down's syndrom

Summary The activities of 21 enzymes of the energy supplying and the glutathione metabolism were estimated in isolated erythrocytes from children with Down's syndrome aged from 1 to 5 1/2 years. All 20 patients were trisomic for chromosome 21. The following enzymes were estimated by means of optic assays: All glycolytic enzymes, two enzymes of the hexosephosphate shunt and two of the citric cycle, NADP- and NAD-dependent glutathione reductase, glutamic-oxaloacetic transaminase, adenylate kinase, and Mg²⁺-activated ATPase.The activities of five enzymes, i.e. phosphofructokinase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, glutathione reductase (NADP), and glutamic-oxaloacetic transaminase, were significantly elevated as compared to age-matched controls. The 50% increase of phosphofructokinase-activity is assumed to provide suggestive evidence for a controlling gene-locus on chromosome 21. The remaining findings are not considered to be directly related to the chromosomal abnormality.

---

---

Direcktor: Prof. Dr. H.-R. Wiedemann

---

Nach einem Vortrag auf der 10. Tagung der Gesellschaft für Anthropologie und Human-genetik, Königstein (Ts.), 22.–25. 10. 1967.

---

Die Untersuchungen wurden durch die dankenswerte Unterstützung der Deutschen Forschungsgemeinschaft ermöglicht.     

Light-Harvesting Complex Protein LHCBM9 Is Critical for Photosystem II Activity and Hydrogen Production in Chlamydomonas reinhardtii

Photosynthetic organisms developed multiple strategies for balancing light-harvesting versus intracellular energy utilization to survive ever-changing environmental conditions. The light-harvesting complex (LHC) protein family is of paramount importance for this function and can form light-harvesting pigment protein complexes. In this work, we describe detailed analyses of the photosystem II (PSII) LHC protein LHCBM9 of the microalga Chlamydomonas reinhardtii in terms of expression kinetics, localization, and function. In contrast to most LHC members described before, LHCBM9 expression was determined to be very low during standard cell cultivation but strongly increased as a response to specific stress conditions, e.g., when nutrient availability was limited. LHCBM9 was localized as part of PSII supercomplexes but was not found in association with photosystem I complexes. Knockdown cell lines with 50 to 70% reduced amounts of LHCBM9 showed reduced photosynthetic activity upon illumination and severe perturbation of hydrogen production activity. Functional analysis, performed on isolated PSII supercomplexes and recombinant LHCBM9 proteins, demonstrated that presence of LHCBM9 resulted in faster chlorophyll fluorescence decay and reduced production of singlet oxygen, indicating upgraded photoprotection. We conclude that LHCBM9 has a special role within the family of LHCII proteins and serves an important protective function during stress conditions by promoting efficient light energy dissipation and stabilizing PSII supercomplexes.     

Multipoint linkage analysis in Menkes disease.

Linkage analyses were performed in 11 families with X-linked Menkes disease. In each family more than one affected patient had been diagnosed. Forty informative meioses were tested using 11 polymorphic DNA markers. From two-point linkage analyses high lod scores are seen for DXS146 (pTAK-8; maximal lod score 3.16 at recombination fraction [theta] = .0), for DXS1 (p-8; maximal lod score 3.44 at theta = .0), for PGK1 (maximal lod score 2.48 at theta = .0), and for DXS3 (p19-2; maximal lod score 2.90 at theta = .0). This indicates linkage to the pericentromeric region. Multilocus linkage analyses of the same data revealed a peak for the location score between DXS146(pTAK-8) and DXYS1X(pDP34). The most likely location is between DXS159 (cpX289) and DXYS1X(pDP34). Odds for this location relative to the second-best-supported region, between DXS146(pTAK-8) and DXS159 (cpX289), are better than 74:1. Visualization of individual recombinant X chromosomes in two of the Menkes families showed the Menkes locus to be situated between DXS159(cpX289) and DXS94(pXG-12). Combination of the present results with the reported absence of Menkes symptoms in male patients with deletions in Xq21 leads to the conclusion that the Menkes locus is proximal to DXSY1X(pDP34) and located in the region Xq12 to Xq13.3.     

A thermosensitive lesion in a Chinese hamster cell mutant causing differential effects on the acidification of endosomes and lysosomes

We previously described the isolation and preliminary characterization of a Chinese hamster ovary cell mutant, termed G.7.1, that carried a temperature-sensitive, conditional-lethal lesion affecting the acidification of vesicles in crude cellular extracts (Marnell, M. H., Mathis, L. S., Stookey, M., Shia, S.-P., Stone, D. K., and Draper, R. K. (1984) J. Cell Biol. 99, 1907-1916). In the present report, we have separated lysosomal vesicles from more buoyant nonlysosomal vesicles by centrifuging cell extracts with Percoll and correlated the acidification defect with nonlysosomal vesicles, including endosomes, but not with secondary lysosomes. Moreover, the acidification of nonlysosomal vesicles prepared from mutant cells grown at the permissive temperature was more sensitive to thermal inactivation than similar vesicles from parental cells, implying that a heat-sensitive component is a normal resident of nonlysosomal vesicles in the mutant. This heat-sensitive component is apparently not associated with lysosomes, or if it is, it does not inhibit lysosomal acidification at the nonpermissive temperature. We also found that the transferrin-mediated uptake of iron is inhibited by 50% in the mutant cells at the nonpermissive temperature and that the inhibition cannot be accounted for by reduced binding or internalization of transferrin.     

Thermodynamic characterization of viral procapsid expansion into a functional capsid shell

The assembly of "complex" DNA viruses such as the herpesviruses and many tailed bacteriophages includes a DNA packaging step where the viral genome is inserted into a preformed procapsid shell. Packaging triggers a remarkable capsid expansion transition that results in thinning of the shell and an increase in capsid volume to accept the full-length genome. This transition is considered irreversible; however, here we demonstrate that the phage λ procapsid can be expanded with urea in vitro and that the transition is fully reversible. This provides an unprecedented opportunity to evaluate the thermodynamic features of this fascinating and essential step in virus assembly. We show that urea-triggered expansion is highly cooperative and strongly temperature dependent. Thermodynamic analysis indicates that the free energy of expansion is influenced by magnesium concentration (3-13?kcal/mol in the presence of 0.2-10?mM Mg(2+)) and that significant hydrophobic surface area is exposed in the expanded shell. Conversely, Mg(2+) drives the expanded shell back to the procapsid conformation in a highly cooperative transition that is also temperature dependent and strongly influenced by urea. We demonstrate that the gpD decoration protein adds to the urea-expanded capsid, presumably at hydrophobic patches exposed at the 3-fold axes of the expanded capsid lattice. The decorated capsid is biologically active and sponsors packaging of the viral genome in vitro. The roles of divalent metal and hydrophobic interactions in controlling packaging-triggered expansion of the procapsid shell are discussed in relation to a general mechanism for DNA-triggered procapsid expansion in the complex double-stranded DNA viruses.