Solution Structure of Histone Chaperone ANP32B: Interaction with Core Histones H3-H4 through Its Acidic Concave Domain

Eukaryotic gene expression is regulated by histone deposition onto and eviction from nucleosomes, which are mediated by several chromatin-modulating factors. Among them, histone chaperones are key factors that facilitate nucleosome assembly. Acidic nuclear phosphoprotein 32B (ANP32B) belongs to the ANP32 family, which shares N-terminal leucine-rich repeats (LRRs) and a C-terminal variable anionic region. The C-terminal region functions as an inhibitor of histone acetylation, but the functional roles of the LRR domain in chromatin regulation have remained elusive. Here, we report that the LRR domain of ANP32B possesses histone chaperone activity and forms a curved structure with a parallel β-sheet on the concave side and mostly helical elements on the convex side. Our analyses revealed that the interaction of ANP32B with the core histones H3-H4 occurs on its concave side, and both the acidic and hydrophobic residues that compose the concave surface are critical for histone binding. These results provide a structural framework for understanding the functional mechanisms of acidic histone chaperones.     

Four hours of daily contact with sexually active males is sufficient to induce fertile ovulation in anestrous goats

The study was conducted on two consecutive years to determine whether ovulatory activity can be induced in anovulatory goats by exposing them to sexually active bucks for 4, 8, 12 or 16 h per day during 15 consecutive days. In experiment 1, females remained continuously in the experimental pens where they were in contact with males. One group remained isolated from males (controls) and four other groups were exposed to sexually active males for 4, 8, 12 or 16 h per day. In experiment 2, females were taken away to “resting” pens free of male odours between the periods of contact with bucks. They were allocated to 5 groups as in experiment 1. Ovulations were determined by progesterone plasma levels and transrectal ultrasonography. Pregnancy was determined by abdominal ultrasonography. In both experiments, more than 90% of females exposed to the bucks had at least one ovulation during the whole experiment whereas only 11 or 0% (experiments 1 and 2, respectively) did so in the control group (P < 0.001). Furthermore, the proportion of females ovulating did not differ among groups depending on duration of contact with bucks (P > 0.05). In both experiments, pregnancy rates were not affected by the daily duration of contact with males (P > 0.05). To conclude, 4 h of daily contact with sexually active males is sufficient to stimulate ovulatory activity in anovulatory goats and this effect is not due to the presence of olfactory cues from the males remaining in the pens.     

Reactivity difference between protolytic forms of some macrocyclic chromium(III) complexes in ligand substitution and electron transfer processes

The review provides insight into the mechanism of ligand substitution and electron transfer (from chromium(III) to iron(III)) by comparison of the reactivity of some tetraazamacrocyclic chromium(III) complexes in the conjugate acid-base forms. Use of two geometrical isomers made possible to estimate the influence of geometry and protolytic reactions in trans and cis position towards the leaving group on the rate enhancement. Studies on the reaction rates in different media demonstrated the role played by outer sphere interactions in a monodentate ligand substitution.     

Synthesis, structure and photocatalytic activity of a remarkably bent, cofacial ethene-linked diiron (III) μ-oxobisporphyrin

Synthesis and characterization of cis and trans 1,2-bis[Chloroiron(III) 5-(2,3,7,8,12,13,17,18-octaethylporphyrinyl)]ethenes are described. Upon treatment with 5% NaOH, cis form of the molecule immediately converts to remarkably bent diiron (III)-μ-oxo bisporphyrin which transforms to cis bisChloroFe(III)porphyrin again by the addition of 5% HCl. This facile transformation is reversible with sharp change in color in which the bisporphyrin platform ‘open’ and ‘close’ its binding pockets with very high vertical flexibility in a single molecular framework. Single crystal X-ray structural characterization reveals cis diiron(III)-μ-oxo bisporphyrin in which Fe-O-Fe unit is remarkable bent with 150.9(2)° angle. Two porphyrin rings in the molecule are not slipped but face-to-face in a fully eclipsed geometry and are placed so close that some of the carbon atoms from each of the macrocycles are driven to be essentially less than the van der Waals contacts (<3.4 Å). Two rings in the oxo-bridged dimer also make the interplanar angle of 27.7° instead of expected angle of 60° due to the bridging alkenic bond. EPR, ¹H NMR and Mössbauer spectral data are indicative of strong anti-ferromagnetic coupling between two high-spin iron(III) centers via bridging oxo group. The complex catalyzes the rapid photoinduced oxygenation of phosphites under mild condition using aerial O₂. Electrochemical data reveals that the diiron(III)-μ-oxo bisporphyrin in dichloromethane undergoes four reversible/quasi-reversible one electron oxidation and one electron reductions. The presence of two porphyrin macrocycles within a short distance in the μ-oxo species makes the porphyrin core highly nonplanar and more electron rich that might responsible for easier oxidations compared to [Fe(OEP)]₂O.     

Deuterium effect on ionization and fragmentation patterns of monosaccharides ionized by atmospheric pressure chemical ionization

Glucose, galactose, and mannose in H₂O and D₂O were ionized by an atmospheric pressure chemical ionization (APCI) method. Isotope effects on fragmentation patterns of the monosaccharides were examined by deuterium replacement of the -OH groups to distinguish the isomers with a single mass spectrometer. The most abundant ions were the [M+H₂O]⁺ and [M_D5+D+D₂O]⁺ for using H₂O and D₂O as solvent and eluent, respectively. Major fragment ions were the [M−OH]⁺ and [M−OH−H₂O]⁺ in H₂O, while those in D₂O were the [M_D5+D−D₂O]⁺ and [M_D5+D−2D₂O]⁺. The differences in the product ions generated in H₂O and D₂O were due to enhancement of the strength of hydrogen bonding by the deuterium replacement. Variations of the ion intensity ratios of the [M−OH]⁺/[M−OH−H₂O]⁺ and [M_D5−OD]⁺/[M_D5−OD−D₂O]⁺ with the fragmentor voltage showed different trends depending on the kind of monosaccharides. By comparing the ion intensity ratios of the [M+H₂O]⁺/M⁺, [M_D5+D+D₂O]⁺/[M_D5+D]⁺, [M−OH]⁺/[M−OH−H₂O]⁺, and [M_D5+D−D₂O]⁺/[M_D5+D−2D₂O]⁺, it was possible to distinguish the isomers of monosaccharides.     

Disulfide linkage in the coiled-coil domain of subunit H of A1AO ATP synthase from Methanocaldococcus jannaschii and the NMR structure of the C-terminal segment H85-104

The C-terminal residues 98-104 are important for structure stability of subunit H of A₁A_O ATP synthases as well as its interaction with subunit A. Here we determined the structure of the segment H_85-104 of H from Methanocaldococcus jannaschii, showing a helix between residues Lys90 to Glu100 and flexible tails at both ends. The helix-helix arrangement in the C-terminus was investigated by exchange of hydrophobic residues to single cysteine in mutants of the entire subunit H (H_I93C, H_L96C and H_L98C). Together with the surface charge distribution of H_85-104, these results shine light into the A-H assembly of this enzyme.     

Olfactory discrimination among sex pheromone stereoisomers: chirality recognition by pink hibiscus mealybug males

Our previous field studies suggested that the two chiral centers in the sex pheromone of pink hibiscus mealybug, Maconellicoccus hirsutus, could elicit different male responses. The chiral center in the acid moiety of the pheromone seemed to be more critical than the alcohol portion of the pheromone molecule for attractiveness. The objective of the current study was to test this hypothesis by deploying stereoisomeric blends in pheromone traps. Captures of male M. hirsutus showed that pheromone with the naturally occurring (R)-maconelliyl (S)-2-methylbutanoate and (R)-lavandulyl (S)-2-methylbutanoate [R-S configuration] was most attractive and that pheromone with the unnatural S-S configuration was less attractive. In addition, the RS-R blend (containing R-R and S-R stereoisomers) yielded captures of male M. hirsutus that were comparable to blank controls, and an inhibitory effect was observed when R-R and S-R were combined with naturally occurring R-S blend. These results suggest a unique chirality recognition mechanism; olfactory discrimination among different pheromone stereoisomers depends upon both asymmetric centers. The S configuration on the acid moiety elicits attraction, whereas the R configuration induces inhibition. However, the attractive activity shows some degree of tolerance toward chirality change in the alcohol portion of the pheromone molecules.     

多效唑对大花高代组培苗生长的效应

以MS0、MS+PP3330.5mg·L-1、MS+PP3331mg·L-1和MS+PP3332mg·L-14种培养基研究多效唑(PP333)对大花高代组培苗生长影响的结果表明,PP333对大花高代组培苗有明显的矮化效应,在0.5￣2mg·L-1PP333范围内,浓度越高,植株越矮,节间越短,叶形指数越小。根据组培苗的真叶和根的发生动态、矮化和黄化程度综合考虑,认为MS+PP3331mg·L-1是较理想的矮化大花高代苗的培养基。     

Barley anther culture: effects of annual cycle and spike position on microspore embryogenesis and albinism

The effect of donor plants annual cycle and anther/spike position on the production of microspore-derived plants and albinism were studied. We used the winter cv. Igri and the spring cv. Cork, known to respond similarly in anther culture but to produce 78% and 2% of green plants, respectively. In both cvs. the number of microspore-derived plants was significantly higher when the anthers were collected from January to July than from August to December. However, during this period the proportion of albino plants was not altered. Conversely, the anther response decreased from 76.6 to 31.5% in Igri and from 58.8 to 32.0% in Cork when the donor spike originates from the main shoot or the fourth tiller. Significantly, anthers collected from spike of the second tiller enabled us to drastically increase the proportion of regenerated green plantlets, by 16% in Igri and 1800% in Cork.     

Interaction between mammalian glyceraldehyde-3-phosphate dehydrogenase and L-lactate dehydrogenase from heart and muscle

The exceptionally high protein concentration in living cells can favor functional protein-protein interactions that can be difficult to detect with purified proteins. In this study we describe specific interactions between mammalian D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and L-lactate dehydrogenase (LDH) isozymes from heart and muscle. We use poly(ethylene-glycol) (PEG)-induced coprecipitation and native agarose electrophoresis as two independent methods uniquely suited to mimic some of the conditions that can favor protein-protein interaction in living cells. We found that GAPDH interacts with heart or muscle isozymes of LDH with approximately one-to-one stoichiometry. The interaction is specific; GAPDH shows interaction with two LDH isozymes that have very different net charge and solubility in PEG solution, while no interaction is observed with GAPDH from other species, other NAD(H) dehydrogenases, or other proteins that have very similar net charge and molecular mass. Analytical ultracentrifugation showed that the LDH and GAPDH complex is insoluble in PEG solution. The interaction is abolished by saturation with NADH, but not by saturation with NAD(+) in correlation with GAPDH solubility in PEG solution. The crystal structures show that GAPDH and LDH isozymes share complementary size, shape, and electric potential surrounding the active sites. The presented results suggest that GAPDH and LDH have a functional interaction that can affect NAD(+)/NADH metabolism and glycolysis in living cells.