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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The formation of compact double-stranded DNA molecules in PEG-containing watersalt solutions (0.3 M NaCl) may be observed within the pH-range 3-10; i.e. under conditions at which parameters of double-stranded DNA helices are not strongly different from those of B-form. At pH less than 3, when regularity of double helices is significantly changed, the formation of the specific compact particles of DNA in PEG-containing solutions does not take place. Denaturation of the compact form of DNA in PEG-containing solution is accompanied by disappearance of the negative band in CD spectrum. Hyperchromic effect of denaturation of DNA compact form is uninformative because of the influence of the light-scattering by compact DNA molecules.
Mol Biol (Mosk)
PMID:[A compact form of DNA in solution. 2. Peculiarities of acidic titration of double-stranded DNA in PEG-containing water-salt solutions]. 0 28

Influence of pH on absorbance and CD-spectra of DNA in PEG-containing water-salt solutions has been studied. The changes in the spectra appeared due to disturbance of the DNA secondary structure upon acidification of the medium proir to or after DNA compactization. If acidification preceeds DNA compactization an intense negative band in the CD spectrum inherent to the compact particles is observed at pH values 7-4. The intensity of the band decreases with an increase of the acidity. The size of the compact particles as evaluated from the dependence of the apparent optical density on the wavelength value remains unchanged (about 1200 A). If the solution is strongly acidified (pH 4.0-2.8) and a considerable disturbance in the DNA secondary structure takes place a negative band in the CD spectrum completely disappears. If one acidifies a solution containing preformed DNA compact particles a decrease of the intensity of the CD negative band starts at lower pH values (less than 2.8). This process is accompanied by an increase of the size of the particles. Acidic "denaturation" of DNA within the compact particles (pH approximately 2.5) is followed by a dissappearance of the CD negative band and a considerable increase of the particle size. The data obtained indicate that the specific arrangement of DNA strands manifested in a CD negative band depends on the defects in the DNA secondary structure.
Mol Biol (Mosk)
PMID:[The compact form of DNA in solution. IV. The effect of secondary structure defectiveness on the arrangement of double-chained DNA molecules into compact particles]. 0 1

The effects of acid on fragmented sarcoplasmic reticulum from rabbit white skeletal muscle have been studied. Brief exposure of sarcoplasmic reticulum membranes to pH values in the range 5.5 to 6.0 at 37 degrees caused rapid inactivation of calcium accumulation measured at 25 degrees in the presence of oxalate (calcium uptake) while (Ca2+, Mg2+)-ATPase (EC 3.6.1.3) activity was enhanced by 75%. ATPase activity, measured at 37 degrees in the absence of oxalate and in the calcium steady state, was unaltered when calcium uptake was inactivated. Calcium efflux from sarcoplasmic reticulum vesicles, previously loaded passibely with 45CaCl2, was only slightly increased when calcium uptake was abolished. At still lower pH values, 5.0 to 5.5, (Ca2+, Mg2+)-ATPase was inactivated while Mg2+ ATPase was more acid-resistant. Acid inactivation of calcium uptake followed simple first order kinetics for at least 80% of the time course. The rate constant, k, increased from 0.043 min-1 to 1.63 min-1 between pH 6.50 and pH 5.35. At pH 4.65, Ea, the energy of activation, was 31 kcal mol-1 between 24 degrees and 43 degrees. Inactivation, once initiated, was irreversible. Aged suspensions of sarcoplasmic reticulum were more sensitive to acid inactivation. Ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid enhanced inactivation, and calcium specifically protected against inactivation with half-maximal effect at 1 to 2 mM. The sulfhydryl reagent, dithiothreitol (1 mM), caused significantly increased rates of inactivation. Calcium binding was studied by dual wavelength spectrophotometry and stopped flow analysis. Acid inactivation distinguished two ATP-induced binding sites, previously described (Entman, M. L., Snow, T. R., Freed, D., and Schwartz, A. (1973) J. Biol. Chem. 248, 7762-7772) as a superficial Mg2+-independent Site A which binds and releases calcium rapidly and a deeper Mg2+-dependent Site B which binds and releases calcium more slowly. Rates of binding to both sites were decreased by acid inactivation. Binding of calcium to Site A increased, however, from 4.6 to 6.4 nmol mg of protein-1 whereas that to Site B decreased from 17.0 to 6.9 nmol mg of protein-1. Passive binding of calcium to sites of medium affinity (K = 7 X 10(4) M-1) was unaffected by acid inactivation of calcium uptake. Temperature dependence of (Ca2+, Mg2+)-ATPase was unchanged in the range 9-34 degrees. Above 34 degrees, the higher activation energy process (Ealpha = 33.7 kcal mol-1) observed in control sarcoplasmic reticulum and thought to arise from a conformational change in the ATPase (Inesi, G., Millman, M., and Eletr, S. (1973) J. Mol. Biol. 81, 483-504) was diminished by acid inactivation (Ealpha = 8.2 kcal mol-1) in a manner suggesting that it is related to active calcium transport. The ATP in equilibrium 32Pi exchange reaction was diminished by acid, but 25% of the activity remained when calcium uptake was completely abolished...
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PMID:Proton inactivation of Ca2+ transport by sarcoplasmic reticulum. 1 42

Paracrystalline fibers of deoxygenated sickle hemoglobin in erythrocytes or concentrated solutions exhibit a phase transformation to a fully crystalline state. X-ray diffraction patterns of the fiber and crystallites are similar except in two respects: the equatorial spacings of the fibers suggest that they pack into a square lattice with a = 220 A, whereas those of the crystals can be indexed on the basis of a net of 187 A by 54 A, and the second-order near-meridional reflections are strong on the fiber pattern but weak on that of the crystallites. The crystallites are isomorphous with single crystals grown in polyethylene glycol solution at pH 4.5 whole structure has been determined at near-atomic resolution (Wishner, B.C., Ward, K.B. Lattmen, E.E. & Lowve, W.E. (1975) J. Mol. Biol. 98, 179-194). Double filaments of molecules with an axial repeat of 64 A comprise the basic unit of both the crystal and fiber structures. Each filament of the pair is translated with respect to its neighbor by half a molecular diameter along the fiber axis. The two filaments are held together by contacts made by Val 6beta in the molecules of one strand with hydrophobic side chains of the molecule in the neighboring strand. This interaction is probably the cause of the aggregation of filaments into fibers that leads to the sickling of erythrocytes.
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PMID:X-ray diffraction studies of fibers and crystals of deoxygenated sickle cell hemoglobin. 3 49

Double-stranded polyribonucleotides (a replicative form of phage f2 RNA--dsRNA and poly(A) poly(U), can adopt a compact from in solutions, containing NaCl and poly(ethylene glycol) (PEG). According to electron-microscopic observations dsRNA compact particles have the form of disks or doughnuts 200--400 A in diameter. X-ray diffraction patterns from dense slurries of dsRNA compact particles show a reflection at a spacing of 35 A, which is indicative of the existance of ordered regions in compact particles. The intense positive CD band, which is characteristic of dsRNA and poly(A) poly(U) compact particles, presumably results from the ordered regions in the particles. Heating of the solution leads to the disappearance of the intense positive CD band, probably as a result of the destruction of the ordered structure of compact particles. Heat or acid denatured dsRNA molecules as well as single-stranded molecules of ribosomal RNA also form large particles in PEG-containing solutions. However, X-ray diffraction patterns from these particles do not show the 35 A reflection and the specific positive band is not present in their CD spectra, which indicates that such particles lack ordered internal structure. It is suggested that similar mechanism of compactization of double-stranded polynucleotides (DNA and RNA) exist, and compact particles may be divided into two families (psi+ and psi-), differing by the secondary structure of double-stranded polynucleotides, which form the particles.
Mol Biol (Mosk)
PMID:[Compact form of DNA in solution. XII. Double-stranded polyribonucleotide compacting in the presence of polyethylene glycol]. 3 51

A highly efficient method for transformation of Bacillus subtilis by plasmid DNA is reported. The procedure, which involves polyethylene glycol-induced DNA uptake by protoplasts and subsequent regeneration of the bacterial cell wall, yields up to 80% transformants with an efficiency of 4 x 10(7) transformants per microgram of supercoiled DNA. Plasmids constructed by in vitro ligation or endonuclease-generated fragments of linear plasmid DNA can also transform PEG-treated protoplasts, but at a lower frequency.
Mol Gen Genet 1979 Jan 05
PMID:High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. 10 88

Removal of Ca2+ from the incubation medium by addition of 2 mM ethylene glycol bis-(beta-aminoethyl ether)-N, N'tetraacetic acid (EGTA) leads to at least 75% inhibition of the luteinizing hormone-releasing hormone (LH-RH)-induced accululation of adenosine 3'5'-monophoshpate (cyclic AMP) in rat anterior pituitary gland in vitro. This inhibitory effect of EGTA is reversed by the addition of Ca2+. A half-maximal effect of Ca2+ on LH-RH--induced cyclic AMP accumulation is observed at 2-5 X 10-5 M free Ca2+. The LH-RH-induced LH and FSH release is completely dependent upon the presence of Ca2+ in the incubation medium, a half-maximal effect being measured at 1-2 X 10-4 M free Ca2+. The basal release release of LH is increased upon Ca2+ removal.
Mol Cell Endocrinol 1975 Jan
PMID:Calcium requirement for stimulation of cyclic AMP accumulation in anterior pituitary gland by LH-RH. 16 1

The formation of compact particles from double-stranded DNA molecules in water-salt solutions containing spermidine was studied. It has been shown that in solutions of low ionic strength (0.01 M NaCl) DNA-spermidine complexes have the form of large particles which scatter UV-light. Electron micrographs show that such complexes formed at certain molar ratios spermidine/DNA may exist both as intermolecular aggregates and as toroidal particles 1500 A in diameter. The CD spectra of solutions containing DNA-spermidine complexes are characterized by the positive band (delta epsilon max = 10) at 265--270 nm. The appearance of the positive CD band may be caused by two factors: interaction between DNA and spermidine may lead to the alteration of the DNA secondary structure "in direction to A-form" or intermolecular aggregation, which may change the initial shape of the CD spectrum. The exclusion of spermidine molecules from DNA-spermidine complexes by Na+ ions in presence of poly(ethylene glycol) which occurs as the ionic strength increases from 0.01 to 0.3 does not lead to decompactization of DNA molecules but is accompained by the appearance of the intense negative CD band at 270 nm.
Mol Biol (Mosk)
PMID:[Formation of the compact form of DNA in solution after reaction with spermidine]. 34 64

Interspecific hybrids produced by polyethylene glycol induced fusion of protoplasts from auxotrophic mutants of Aspergillus nidulans and Aspergillus rugulosus were grown in the presence of the recombinogens benomyl and chloral hydrate to stimulate segregation. The A. nidulans parental strains used had a known genetic marker in each linkage group. Hybrids grown on complete medium containing benomyl yielded more segregants. Analysis of the segregants showed that the distribution of A. nidulans linkage groups was random. No specific linkage group appeared in all the segregants. The two parents are closely related taxonomically and the findings from these experiments suggest that a high degree of chromosomal homology may exist between them.
Mol Gen Genet 1979 Feb 26
PMID:Induced segregation in interspecific hybrids of Aspergillus nidulans and Aspergillus rugulosus obtained by protoplast fusion. 37 62

Fusion of protoplasts prepared from haploid strains of Saccharomyces yeasts having identical mating type was induced with the aid of polyethylene glycol. Stable fusion products were isolated by complementation of the auxotrophic markers. Of 64 isolates derived by protoplast fusion between two different haploid strains having alpha mating type, 35 fusion products were estimated from their cell volumes to be diploid, 13 to be triploid and 16 to be tetraploid. The isolates showing tetraploid cell size were thought to have resulted from fusion of three protoplasts of one strain and one protoplast of the other (three-to-one fusion) or from two-to-two fusion. In protoplast fusion of three different haploid strains having alpha mating type, all four possible phenotypes of fusion product were recovered. Fusion products of three different protoplasts were obtained in much lower frequency (2.1 x 10(-6)) than those of two different protoplasts (1.2 x 10(-5) to 1.4 x 10(-4)) in the three other combinations. Genetic analyses revealed that triploid fusion products were formed by protoplast fusion of two different strains as well as of three different strains.
Mol Gen Genet 1979 Jun 20
PMID:Multiple fusion of protoplasts in Saccharomyces yeasts. 38 49


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