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21.
Mlh1-Mlh3, a Meiotic Crossover and DNA Mismatch Repair Factor,Is a Msh2-Msh3-stimulated Endonuclease
Maria V. Rogacheva Carol M. Manhart Cheng Chen Alba Guarne Jennifer Surtees Eric Alani 《The Journal of biological chemistry》2014,289(9):5664-5673
Crossing over between homologous chromosomes is initiated in meiotic prophase in most sexually reproducing organisms by the appearance of programmed double strand breaks throughout the genome. In Saccharomyces cerevisiae the double-strand breaks are resected to form three prime single-strand tails that primarily invade complementary sequences in unbroken homologs. These invasion intermediates are converted into double Holliday junctions and then resolved into crossovers that facilitate homolog segregation during Meiosis I. Work in yeast suggests that Msh4-Msh5 stabilizes invasion intermediates and double Holliday junctions, which are resolved into crossovers in steps requiring Sgs1 helicase, Exo1, and a putative endonuclease activity encoded by the DNA mismatch repair factor Mlh1-Mlh3. We purified Mlh1-Mlh3 and showed that it is a metal-dependent and Msh2-Msh3-stimulated endonuclease that makes single-strand breaks in supercoiled DNA. These observations support a direct role for an Mlh1-Mlh3 endonuclease activity in resolving recombination intermediates and in DNA mismatch repair. 相似文献
22.
ON Rogacheva BF Shchegolev VE Stefanov EV Savvateeva-Popova 《Biochemistry. Biokhimii?a》2012,77(5):465-468
Protein-ligand docking and ab initio calculations have shown that the 3′:5′-AMP phosphorothioate analog (Rp-3′:5′-AMPS) blocks the A326 amide group displacement
typical of transition from the H- to B-conformation within the B-domain of protein kinase A Iα R-subunit. This behavior of
Rp-3′:5′-AMPS leads to the inhibition of initial stages of hydrophobic relay operation. In accordance with the proposed hypothesis,
Rp-3′:5′-AMPS similarly to 3′:5′-AMP forms a hydrogen bond with the amide group of A326; however, the properties of this bond
together with the position of the sulfur atom prevent the movement of A326. Finally, the Rp-3′:5′-AMPS-bound domain appears
to be locked in the H-conformation, which is in agreement with the X-ray data. 相似文献
23.
DNA mismatch repair (MMR) models have proposed that MSH (MutS homolog) proteins identify DNA polymerase errors while interacting with the DNA replication fork. MLH (MutL homolog) proteins (primarily Mlh1-Pms1 in baker's yeast) then survey the genome for lesion-bound MSH proteins. The resulting MSH-MLH complex formed at a DNA lesion initiates downstream steps in repair. MLH proteins act as dimers and contain long (20-30nm) unstructured arms that connect two terminal globular domains. These arms can vary between 100 and 300 amino acids in length, are highly divergent between organisms, and are resistant to amino acid substitutions. To test the roles of the linker arms in MMR, we engineered a protease cleavage site into the Mlh1 linker arm domain of baker's yeast Mlh1-Pms1. Cleavage of the Mlh1 linker arm in vitro resulted in a defect in Mlh1-Pms1 DNA binding activity, and in vivo proteolytic cleavage resulted in a complete defect in MMR. We then generated a series of truncation mutants bearing Mlh1 and Pms1 linker arms of varying lengths. This work revealed that MMR is greatly compromised when portions of the Mlh1 linker are removed, whereas repair is less sensitive to truncation of the Pms1 linker arm. Purified complexes containing truncations in Mlh1 and Pms1 linker arms were analyzed and found to have differential defects in DNA binding that also correlated with the ability to form a ternary complex with Msh2-Msh6 and mismatch DNA. These observations are consistent with the unstructured linker domains of MLH proteins providing distinct interactions with DNA during MMR. 相似文献
24.
V G Zilov A P Patyshakuliev S K Rogacheva L I Ivanova 《Biulleten' eksperimental'no? biologii i meditsiny》1992,114(11):455-457
An ability of substance P (15 nmol intracerebroventricularly) to transform functional properties of the hypothalamic feeding center manifested in escape reaction in response to threshold electrical stimulation of this motivation center in rabbits. Beta-adrenergic antagonists inderal and obsidan (0.25, 0.5 mg, respectively) injected intravenously were found to restore feeding elicited by electrostimulation of the hypothalamic feeding center. However, escape reaction, elicited from the feeding center under substance P was found unchanged after intravenous administration of 0.25 mg kalipsol. The data obtained suggested an important role of beta-adrenergic mechanisms in substance P transformation of the functional properties of the hypothalamic feeding centre. 相似文献
25.
Rogacheva SM Denisova SA Shul'gin SV Somov AIu Kuznetsov PE 《Radiatsionnaia biologiia, radioecologiia / Rossi?skaia akademiia nauk》2008,48(4):474-480
The study of the action of the electromagnetic radiation (EMR) of low intensity (10 microW/cm2) in the range of frequencies 120-170 GHz at the test-reaction of Infusoria Paramecium caudatum was carried out. The resonant character of the effects was established. The EMR action at 156.6 and 161.3 GHz caused the increase of infusorians mobility, the action at frequencies 151.8, 155.7, 167.1 GHz caused the mobility reduction. Isolated and combined with EMR effects of nicotine (10(-4)-10(-15) mol/l) and antimicrobial drug metronidazole (10(-5), 10(-8), 10(-9) mol/l) were investigated. The radiation at the frequency 167.1 GHz was shown to reduce the effect of nicotine (10(-9) mol/l) and to enhance the effect of metronidazole (10(-9) mol/l). This phenomenon may be explained by different effects of the substances in low concentration at the water hydrogen bonds net structure. 相似文献
26.
S A Rogacheva V M Luzanov E N Kirillova A E Baranov K N Muksinova 《Radiobiologiia》1990,30(6):769-773
A human recombinant granulocytic-and-macrophagic colony-stimulating factor (rGM-CSF) administered repeatedly to irradiated (10 Gy) CBA mice increased CFUs and CFU-GM content, the number of bone marrow granulocytes and erythronormoblasts, and spleen and peripheral blood cellularity. The survival rate of exposed (9.7 Gy) mice repeatedly injected with rGM-CSF increased from 25% (control) to 90%. 相似文献
27.
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28.
The respiratory activity of Rhodococcus rhodochrousM8 cells containing nitrile hydratase and amidase was studied in the presence of nitriles and amides of carbonic acids. The culturing of cells with acrylonitrile and acrylamide yielding maximum respiratory activity was studied. The optimum conditions for measurements and maintenance of respiratory activity were found. Curves for the linear concentration dependence of cell respiratory activity on 0.01–0.5 mM acrylonitrile, 0.025–1.0 mM acetonitrile, and 0.01–0.1 mM acrylamide were plotted. The selectivity of cell respiratory activity for some substrates was analyzed. 相似文献
29.
V G Zilov S K Rogacheva L I Ivanova 《Biulleten' eksperimental'no? biologii i meditsiny》1983,96(11):74-77
Chronic experiments on rabbits with electrodes implanted into different limbic-midbrain structures were made to study the effects of a single intravenous injection of ethanol (0.5 g/kg) on the background EEG during formation of food motivation and avoidance behavior from the criterion of the power of the main EEG rhythms. Intravenous injection of ethanol resulted in an increase in the power of beta-, alpha- and theta-rhythms in the frontal cortex, and in that of alpha- and theta-rhythms in the occipital area of the neocortex. New patterns of the powers of the main EEG rhythms recorded in animals exposed to ethanol during electric stimulation of the lateral and ventromedial hypothalamus, evoking food motivation and avoidance behavior, as well as during electrical stimulation of the dorsal hippocamp and mesencephalic reticular formation that correlate with changes in the functions of the study limbic-mesencephalic structures attest to profound ethanol-induced abnormalities of the central mechanisms of food motivation and avoidance behavior. 相似文献
30.
ANNA MARTINEZ EL MOUKHTAR ALIOUAT MURIEL POTTIER NAUSICÂA GANTOIS CLAIRE PINÇON ANNIE STANDAERT-VITSE EDUARDO DEI-CAS CÉCILE-MARIE ALIOUAT-DENIS 《The Journal of eukaryotic microbiology》2009,56(5):446-453
ABSTRACT. The separation of Pneumocystis carinii life-cycle stages while preserving infectivity is a hitherto unresolved challenge. We describe an original, reproducible, and efficient method for separating trophic from cystic forms of P. carinii using a high-speed cell sorter. The large amounts of highly purified (99.6±0.3%) infectious trophic and cystic forms can now be used to elucidate the poorly understood P. carinii life cycle. 相似文献