Sequence Determinants of GLUT1 Oligomerization: ANALYSIS BY HOMOLOGY-SCANNING MUTAGENESIS*

The human blood-brain barrier glucose transport protein (GLUT1) forms homodimers and homotetramers in detergent micelles and in cell membranes, where the GLUT1 oligomeric state determines GLUT1 transport behavior. GLUT1 and the neuronal glucose transporter GLUT3 do not form heterocomplexes in human embryonic kidney 293 (HEK293) cells as judged by co-immunoprecipitation assays. Using homology-scanning mutagenesis in which GLUT1 domains are substituted with equivalent GLUT3 domains and vice versa, we show that GLUT1 transmembrane helix 9 (TM9) is necessary for optimal association of GLUT1-GLUT3 chimeras with parental GLUT1 in HEK cells. GLUT1 TMs 2, 5, 8, and 11 also contribute to a less abundant heterocomplex. Cell surface GLUT1 and GLUT3 containing GLUT1 TM9 are 4-fold more catalytically active than GLUT3 and GLUT1 containing GLUT3 TM9. GLUT1 and GLUT3 display allosteric transport behavior. Size exclusion chromatography of detergent solubilized, purified GLUT1 resolves GLUT1/lipid/detergent micelles as 6- and 10-nm Stokes radius particles, which correspond to GLUT1 dimers and tetramers, respectively. Studies with GLUTs expressed in and solubilized from HEK cells show that HEK cell GLUT1 resolves as 6- and 10-nm Stokes radius particles, whereas GLUT3 resolves as a 6-nm particle. Substitution of GLUT3 TM9 with GLUT1 TM9 causes chimeric GLUT3 to resolve as 6- and 10-nm Stokes radius particles. Substitution of GLUT1 TM9 with GLUT3 TM9 causes chimeric GLUT1 to resolve as a mixture of 6- and 4-nm particles. We discuss these findings in the context of determinants of GLUT oligomeric structure and transport function.     

APOLAR EMBRYOS OF FUCUS RESULTING FROM OSMOTIC AND CHEMICAL TREATMENT

Embryos of the brown alga Fucus vesiculosas L. were grown as populations in glass petri dishes in seawater at 15 C in continuous low-intensity unilateral fluorescent illumination for periods up to 2 weeks. A quantitative estimate of increase in nuclear number was made from acetocarmine squash preparations of samples taken at 12-or-24 hr intervals. Over the period of 2-6 days embryos showed a doubling time of about 12-18 hr. Under normal seawater culture conditions each embryo formed a single rhizoid. When grown in seawater supplemented with sugar concentrations above 0.4 m , Fucus embryos developed as multicellular spherical embryos lacking rhizoids. In 0.6 m sucrose-seawater, 97% of the embryos were apolar at 2 days; only 37% were apolar at 4 days, many having recovered from the sucrose inhibition. Some embryos remained apolar after growth in 0.6 m sucrose for 2 weeks. Nuclear counts showed that sucrose-seawater markedly inhibited the rate of cell division. Other sugars including D-glucose, D-fructose, D-galactose and the sugar alcohol D-mannitol were also effective. When apolar embryos grown in sucrose-seawater were returned to seawater, embryo growth resumed at the normal seawater rate, judged from nuclear counts. Such embryos formed multiple rhizoids, varying from two to eight rhizoids per embryo, which developed on the embryo quadrant or half away from the unilateral light. Each of the multiple rhizoids originated from a single small cell in the periphery of the multicellular spherica embryo. Thus the rhizoid-forming stimulus apparently had been subdivided among a number of the cells of the apolar embryos. The implications of this finding are discussed. Attempts to produce multiple rhizoids by treatment of embryos with indoleacetic acid or 2,4-dichlorophen-oxyacetic acid failed. However, embryos treated with 10⁻⁴ M or 5 × 10⁻⁵ m 2,3,5-triiodobenzoic acid formed 40 and 30% multiple rhizoids, respectively, suggesting that some chemical, perhaps hormonal, mechanism is involved in polarization and rhizoid initiation in Fucus embryogenesis.     

Ribosomal protein S12 as a site for streptomycin resistance in Nicotiana chloroplasts

Summary A streptomycin resistant Nicotiana plastome mutant, X/str ^R6, was subjected to molecular analysis. In this mutant, a single nucleotide transition, C » T, in the chloroplast gene for ribosomal protein S12 alters codon 90 from proline to serine while the nucleotide sequence of the chloroplast 16 S rRNA gene is identical to that of the wild type. Mutant X/str ^R6 thus differs from several previously reported streptomycin resistant chloroplast mutants which are altered in the gene for 16 S rRNA.     

Complementation in Nonconidiating Mutants of Trichoderma

Nonconidiating (Con(-)) mutants were isolated from wild-type and color mutants of the fungus Trichoderma viride Pers. ex Fries. Heterokaryons were easily produced and maintained, and the complementation relationships among the Con(-) mutants were established. Most Con(-) mutants could complement one or more of the other Con(-) mutants. When marked Con(-) mutants were mixed with marked Con(+) testers, conidiating heterokaryons were formed. The conidia thus obtained produced only the parental type colonies after replating, indicating that nonconidiation is a nuclear characteristic. Allowing two Con(-) colonies to meet and produce a heterokaryon, it was found that the migration of nuclei reached a rate of 5 mm per hr, which is several times greater than the rate of hyphal elongation; it was also found that heterokaryosis of a mycelial region preceded its ability to conidiate.     

Study of the inheritance of sex expression in the cucumber. The interaction of major genes with modifying genetic and non-genetic factors

Summary Two major genes and a complex of polygenes affecting sex expression inCucumis sativus L. as well as their interaction with some nongenetic factors were investigated in the present study. The genetic factorst was found to affect sex by inducing a shift of the predetermined flowering pattern of this plant, in the direction of its base. As this pattern is composed of a staminate stage followed by a mixed (staminate-pistillate) stage and a pistillate stage, a double dose ofst will induce a change from the normal monoecious sex expression (to absolute gynoecism.The second major gene studied,m, known previously to control sex in the individual flower (m/m—andromonoecious,M—monoecious), also interacts with factors affecting the flowering pattern by inducing male tendency.Using an ordinary commercial monoecious stock additional modifying factors for sex expression were demonstrated, by eight generations of selection for high (in male direction) and for low (in female direction) node number to the first pistillate flower. A comparison of the means and frequency distributions of node number of the two selected lines, their F₁ and F₂, indicated polygenic control of this character.It was found that two non-genetic factors, day length and gibberellic acid (GA), may mimic the genetic factors for sex expression. Furthermore, evidence was presented, indicating that this sex controlling ability of GA and the modifying genes may be based on physiological conditions common to those two genetic and nongenetic factors.The overall mechanism of sex control in the cucumber and in other plants was discussed and a hypothetic model for the evolution of dioecism in flowering plants was proposed.     

Photoluminescence and thermoluminescence properties of pure and rare-earth-doped ZrO2 prepared by Pechini-type sol–gel

In this article, photoluminescence (PL) and thermoluminescence (TL) properties of ZrO₂, ZrO₂:Dy³⁺, ZrO₂:Dy³⁺–Gd³⁺, ZrO₂:Dy³⁺–Yb³⁺, ZrO₂:Dy³⁺–Er³⁺, and ZrO₂:Dy³⁺–Sm³⁺ phosphors synthesized by the Pechini method were investigated. The crystal structure, thermal properties, morphology, PL and TL properties were investigated using X-ray powder diffraction (XRD), differential thermal analysis/thermogravimetric analysis (DTA/TGA), scanning electron microscopy (SEM), PL and TL, respectively. The room temperature emission bands corresponding to ⁴F_9/2 → ⁶H_J (J = 9/2, 11/2, 13/2 and 15/2) transitions of Dy³⁺ ions were measured. The phosphors were analysed using T_m–T_STOP, variable dose, and computerized glow curve fitting methods. Reusability, dose–response, and fading characteristics were investigated. The phosphors have a natural TL emission that vanished by heating treatment. Moreover, new peaks with similar properties to the natural emissions were observed after high-dose irradiation and long-term fading experiments. The glow curves of the phosphors have 13 individual peaks and many low- and high-temperature satellite peaks. The origin of the peaks is ZrO₂ host material and doping with rare-earth ions (Gd³⁺, Dy³⁺, Yb³⁺, Er³⁺ and Sm³⁺) does not lead to a new glow peak. The dopants cause drastic changes in individual peak intensities of ZrO₂.The initial fading rates of all the phosphors are relatively fast, but they slow down as time goes on.     

Comparative bioinformatics analysis and abiotic stress responses of expansin proteins in Cucurbitaceae members: watermelon and melon

Protoplasma - Watermelon and melon are members of the Cucurbitaceae family including economically significant crops in the world. The expansin protein family, which is one of the members of the...     

Aluminum Toxicity: A Case Study on Tobacco (<i>Nicotiana tabacum</i> L.)

Aluminum is an abundant metal in the earth’s crust that turns out to be toxic in acidic environments. Many plants are affected by the presence of aluminum at the whole plant level, at the organ level, and at the cellular level. Tobacco as a cash crop (Nicotiana tabacum L.) is a widely cultivated plant worldwide and is also a good model organism for research. Although there are many articles on Al-phytotoxicity in the literature, reviews on a single species that are economically and scientifically important are limited. In this article, we not only provide the biology associated with tobacco Al-toxicity, but also some essential information regarding the effects of this metal on other plant species (even animals). This review provides information on aluminum localization and uptake process by different staining techniques, as well as the effects of its toxicity at different compartment levels and the physiological consequences derived from them. In addition, molecular studies in recent years have reported specific responses to Al toxicity, such as overexpression of various protective proteins. Besides, this review discusses data on various organelle-based responses, cell death, and other mechanisms, data on tobacco plants and other kingdoms relevant to these studies.