ВЛИЯНИЕ УСЛОВИЙ ВЫРАЩИВАНИЯ И ЛЕГИРОВАНИЯ ДОНОРНЫМИ ПРИМЕСЯМИ НА МЕХАНИЗМ ПРОВОДИМОСТИ И СПЕКТРЫГЛУБОКИХ УРОВНЕЙ В КРИСТАЛЛАХ TlBr

Studies of electrical characteristics, deep traps spectra, microcathodoluminescence (MCL) spectra of undoped and donor (Pb, Ca) doped TlBr crystals as influenced by growth conditions (Br pressure, Ar pressure, growth in air) are presented. It is shown that, for the 85−320 K temperature range, the crystal conductivity was determined not by ionic conductance but by the density of electrons and holes supplied by the ionization of deep centers. Centers with activation energies of 1−1.2 eV that pin the Fermi level in donor doped crystals are shown to play a prominent role in the recombination of nonequilibrium charge carriers. In undoped crystals the Fermi level is pinned near Ev+0.8 eV and these centers are also active in the recombination of charge carriers and are responsible for the MCL band peak near 1.85 eV. The temperature dependence of photocurrent in undoped crystals is strongly influenced by electron trapping on relatively shallow centers located 0.1−0.2 eV below the conduction band edge. Deep traps spectra revealed the presence of centers with activation energies 0.36, 0.45, 0.6 eV whose concentration increases with donor doping. Doping with Pb or Ca increases the dark resistivity of the crystals by about an order of magnitude, but Pb doping enhances the density of deep traps, which is not favorable for use of this material in radiation detectors.Исследованы электрические характеристики, спектры глубоких ловушек, спектры микрокатодолюминесценции (МКЛ) нелегированных и легированных донорами (Pb, Ca) кристаллов TlBr и изучено влияние на эти характеристики условий выращивания (противодавление брома, противодавление аргона, выращивание на воздухе). Показано, что в исследованном интервале температур (85—320 К) проводимость кристаллов определяется концентрацией электронов и дырок в разрешенных зонах, а не ионной проводимостью. В процессах рекомбинации неравновесных носителей основную роль играют центры с энергией активации 1,0—1,2 эВ, на которых закреплен уровень Ферми в легированных донорами кристаллах. В нелегированных кристаллах уровень Ферми закреплен на центрах с уровнем около Ev+0,8 эВ, которые также участвуют в рекомбинации и ответственны за полосу МКЛ с энергией 1,85 эВ. В температурных зависимостях фототока нелегированных кристаллов большую роль играет прилипание электронов на мелких электронных ловушках с энергией 0,1—0,2 эВ и на более глубоких электронных ловушках. В спектрах глубоких центров обнаружены ловушки с энергиями 0,36, 0,45 и 0,6 эВ, концентрация которых растет при легировании донорами. Легирование Pb или Ca позволяет на порядок повысить удельное сопротивление материала, но легирование Pb приводит к большей концентрации глубоких ловушек, что неблагоприятно для использования материала в радиационных детекторах

Phosphorylation of a chloroplast RNA-binding protein changes its affinity to RNA.

An RNA-binding protein of 28 kDa (28RNP) was previously isolated from spinach chloroplasts and found to be required for 3' end-processing of chloroplast mRNAs. The amino acid sequence of 28RNP revealed two approximately 80 amino-acid RNA-binding domains, as well as an acidic- and glycine-rich amino terminal domain. Upon analysis of the RNA-binding properties of the 'native' 28RNP in comparison to the recombinant bacterial expressed protein, differences were detected in the affinity to some chloroplastic 3' end RNAs. It was suggested that post-translational modification can modulate the affinity of the 28RNP in the chloroplast to different RNAs. In order to determine if phosphorylation accounts for this post-translational modification, we examined if the 28RNP is a phosphoprotein and if it can serve as a substrate for protein kinases. It was found that the 28RNP was phosphorylated when intact chloroplasts were metabolically labeled with [32P] orthophosphate, and that recombinant 28RNP served as an excellent substrate in vitro for protein kinase isolated from spinach chloroplasts or recombinant alpha subunit of maize casein kinase II. The 28RNP was apparently phosphorylated at one site located in the acidic domain at the N-terminus of the protein. Site-directed mutagenesis of the serines in that region revealed that the phosphorylation of the protein was eliminated when serine number 22 from the N-terminus was changed to tryptophan. RNA-binding analysis of the phosphorylated 28RNP revealed that the affinity of the phosphorylated protein was reduced approximately 3-4-fold in comparison to the non-phosphorylated protein. Therefore, phosphorylation of the 28RNP modulates its affinity to RNA and may play a significant role in its biological function in the chloroplast

RNA-binding activities of the different domains of a spinach chloroplast ribonucleoprotein.

An RNA-binding protein of 28 kD (28RNP) has been previously isolated from spinach chloroplasts and was found to be required for 3' end processing of chloroplast mRNAs. The amino acid sequence of 28RNP revealed two approximately 80 amino-acid RNA-binding domains, as well as an acidic and glycine-rich amino terminal domain. Each domain by itself, as well as in combination with other domains, was expressed in bacterial cells and the polypeptides were purified to homogeneity. We have investigated the RNA-binding properties of the different structural domains using UV-crosslinking, saturation binding and competition between the different domains on RNA-binding. It was found that the acidic domain does not bind RNA, but that each of the RNA-binding domains, expressed either individually or together, do bind RNA, although with differing affinities. When either the first or second RNA-binding domain was coupled to the acidic domain, the affinity for RNA was greatly reduced. However, the acidic domain has a positive effect on the binding of the full-length protein to RNA, because the mature protein binds RNA with a better affinity than the truncated protein which lacks the acidic domain. In addition, it was found that a stretch of two or three G residues is enough to mediate binding of the 28RNP, whereas four U residues were insufficient. The implications of the RNA-binding properties of 28RNP to its possible function in the processing of chloroplast RNA is discussed

Polynucleotide Phosphorylase Functions as Both an Exonuclease and a Poly(A) Polymerase in Spinach Chloroplasts

The molecular mechanism of mRNA degradation in the chloroplast consists of sequential events including endonucleolytic cleavage, the addition of poly(A)-rich sequences to the endonucleolytic cleavage products, and exonucleolytic degradation by polynucleotide phosphorylase (PNPase). In Escherichia coli, polyadenylation is performed mainly by poly(A)-polymerase (PAP) I or by PNPase in its absence. While trying to purify the chloroplast PAP by following in vitro polyadenylation activity, it was found to copurify with PNPase and indeed could not be separated from it. Purified PNPase was able to polyadenylate RNA molecules with an activity similar to that of lysed chloroplasts. Both activities use ADP much more effectively than ATP and are inhibited by stem-loop structures. The activity of PNPase was directed to RNA degradation or polymerization by manipulating physiologically relevant concentrations of P(i) and ADP. As expected of a phosphorylase, P(i) enhanced degradation, whereas ADP inhibited degradation and enhanced polymerization. In addition, searching the complete Arabidopsis genome revealed several putative PAPs, none of which were preceded by a typical chloroplast transit peptide. These results suggest that there is no enzyme similar to E. coli PAP I in spinach chloroplasts and that polyadenylation and exonucleolytic degradation of RNA in spinach chloroplasts are performed by one enzyme, PNPase