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Suraj Adhikary Marilyn C. Cato Kriston L. McGary Antonis Rokas Brandt F. Eichman 《DNA Repair》2013,12(3):196-204
Schizosaccharomyces pombe contains two paralogous proteins, Mag1 and Mag2, related to the helix-hairpin-helix (HhH) superfamily of alkylpurine DNA glycosylases from yeast and bacteria. Phylogenetic analysis of related proteins from four Schizosaccharomyces and other fungal species shows that the Mag1/Mag2 duplication is unique to the genus Schizosaccharomyces and most likely occurred in its ancestor. Mag1 excises N3- and N7-alkylguanines and 1,N6-ethenoadenine from DNA, whereas Mag2 has been reported to have no detectible alkylpurine base excision activity despite high sequence and active site similarity to Mag1. To understand this discrepancy we determined the crystal structure of Mag2 bound to abasic DNA and compared it to our previously determined Mag1–DNA structure. In contrast to Mag1, Mag2 does not flip the abasic moiety into the active site or stabilize the DNA strand 5′ to the lesion, suggesting that it is incapable of forming a catalytically competent protein–DNA complex. Subtle differences in Mag1 and Mag2 interactions with the DNA duplex illustrate how Mag2 can stall at damage sites without fully engaging the lesion. We tested our structural predictions by mutational analysis of base excision and found a single amino acid responsible at least in part for Mag2's lack of activity. Substitution of Mag2 Asp56, which caps the helix at the base of the DNA intercalation loop, with the corresponding serine residue in Mag1 endows Mag2 with ?A excision activity comparable to Mag1. This work provides novel insight into the chemical and physical determinants by which the HhH glycosylases engage DNA in a catalytically productive manner. 相似文献
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In this mini-review we present an environmental iron mobility/transport scheme consisting of inter-related controls, whereby the first coordination shell of iron modulates the iron redox potential (E1/2), and the oxidation state of iron controls the chemistry of the first coordination sphere and therefore the immediate chemical environment of the iron. Siderophores (microbially generated iron specific chelators) may be viewed as iron redox mediators. Siderophore chelation of environmental iron in a reduced (Fe(II)) oxidation state results in facile air oxidation of iron due to the negative redox potentials observed for Fe-siderophore complexes. This solubilizes the iron and locks it into a specific coordination environment, thereby preventing hydrolysis and precipitation. The high-spin Fe3+ → Fe+ electron transfer process may be viewed as a switch that controls the thermodynamic stability and kinetic lability of the first coordination shell. Reduction of iron(III)-siderophore complexes to iron(II)-siderophore complexes decreases thermodynamic stability, increases the rate of siderophore ligand exchange, and increases the ease of siderophore donor atom protonation, thus facilitating a rapid turnover of the first coordination shell. Results are presented for iron-siderophore pH and oxidation state dependent speciation studies that are relevant to environmental and microbial iron mobility and transport. 相似文献
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The hypothalamo-hypophysial vascular relationship and intra-hypophysial vasculatisation have been described in order to understand the regulatory mechanism of hypothalamic control over the functions of the pituitary gland. In Glossogobius giuris, the disposition of the blood vessels in the head region is on typical teleostean pattern with certain modifications. The nucleus preopticus is supplied through the nucleus preopticus artery, a small blood vessel arising from the anterior branch of the posterior cerebral artery, whereas the pituitary gland receives blood through a pair of hypophysial arteries. The blood from the pituitary is drained off by the pituitary veins whch pour their blood into the supra-orbital sinus. The anterior cerebral vein after taking the blood from anterior part of the brain including the hypothalamus and the nucleus preopticus joins with the supra-orbital sinus. The hypothalamo-hypophysial portal system is absent in this fish. The saccus vasculosus receives blood from the posterior cerebral artery through a small blood vessel and is collected by a prominent saccus vasculosus vein which pours blood into the supra-orbital sinus before it joins the infra-orbital sinus to form the heat vein. There seems to be no physological connection between the saccus vasculosus and pituitary gland. The highly vascularised neurohypophysis interdigitate with the pars intermedia and extends upto the proximal pars distalis. The blood vessels are restricted to the neurohypophysial extensions only. However, in the rostral pars distalis the blood vessels are present but the neurohypophysis does not extend to this part. The blood capillaries enter the rostral pars distalis from the capillary network on the surface of pituitary gland along with the connected tissue covering of the pituitary. The neurohypophysis shows a greater vascularisation in comparison to that of the other glandular part of the pituitary gland. In the present study of Glossogobius giuris, though an extensive ramification of neurohypophysis occurs with the pars intermedia and the proximal pars distalis, the neurosecretory axons do not innervate the endocrine cells of the pituitary gland and the blood vessels are found restricted to the neurohypophysial extensions except that of the rostral pars distalis. The neuro-vascular way of hypothalamic control over the functions of the pituitary gland seems to be justified as the neurosecretory fibres have been found associated with the blood vessels. 相似文献
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The serum levels of progesterone (Δ4P), 20α-dihydroxyprogesterone (20α-DHP), pregnenolone (Δ5P) and estrogens at different time intervals during Silastic-PVP-PGF2α tube induced midterm abortion were measured by radioimmunoassay in the rat and rabbit. The concentrations of 4P and Δ5P were significantly reduced but that of 20α-DHP was increased significantly as early as 20 hours after PGF2α treatment. Estrogen levels showed slight reduction in the rat but remained unchanged in the rabbit. The ratio of Δ4P/20α-DHP recorded were 2.15 and 16.86 (pretreatment) and dropped to 0.08 and 0.37 (60 hours after insertion of Silastic-PVP-PGF2α tube) in the rat and rabbit, respectively. These findings suggest that the suppression of circulating Δ4P by PGF2α might be the result of a reduced precursor (Δ5P) pool size as well as changes in the enzymic systems responsible for the formation and conversion of Δ4P. It is still to be determined if changes in estrogen levels in the rat have any bearing on abortion induced by PGF2α. 相似文献