Removal of nucleic acids from yeast nucleoprotein complexes by sulfitolysis

1986 ◽  
Vol 34 (1) ◽  
pp. 26-30 ◽  
Author(s):  
Srinivasan Damodaran
Keyword(s):  
Nucleic Acids ◽  
Nucleoprotein Complexes

Snapshot blotting: The transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to EM grids

1992 ◽  
Vol 50 (1) ◽  
pp. 762-763
Author(s):  
Stephen D. Jett
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Nucleic Acid Binding ◽  
Mobility Shift ◽  
Carbon Coated ◽  
Nucleoprotein Complexes ◽  
Mobility Shift Assay ◽  
Protein Nucleic Acid ◽  
Gel Mobility Shift ◽  
Nucleic Acid Interactions

The electrophoresis gel mobility shift assay is a popular method for the study of protein-nucleic acid interactions. The binding of proteins to DNA is characterized by a reduction in the electrophoretic mobility of the nucleic acid. Binding affinity, stoichiometry, and kinetics can be obtained from such assays; however, it is often desirable to image the various species in the gel bands using TEM. Present methods for isolation of nucleoproteins from gel bands are inefficient and often destroy the native structure of the complexes. We have developed a technique, called “snapshot blotting,” by which nucleic acids and nucleoprotein complexes in electrophoresis gels can be electrophoretically transferred directly onto carbon-coated grids for TEM imaging.

scholarly journals Helicase Mechanisms During Homologous Recombination inSaccharomyces cerevisiae

2019 ◽  
Vol 48 (1) ◽  
pp. 255-273 ◽  
Author(s):  
J. Brooks Crickard ◽  
Eric C. Greene
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nucleic Acid ◽  
Homologous Recombination ◽  
Nucleic Acids ◽  
Acid Metabolism ◽  
Model Organism ◽  
Nucleic Acid Metabolism ◽  
Repair Pathway ◽  
Unidirectional Movement ◽  
Nucleoprotein Complexes

Helicases are enzymes that move, manage, and manipulate nucleic acids. They can be subdivided into six super families and are required for all aspects of nucleic acid metabolism. In general, all helicases function by converting the chemical energy stored in the bond between the gamma and beta phosphates of adenosine triphosphate into mechanical work, which results in the unidirectional movement of the helicase protein along one strand of a nucleic acid. The results of this translocation activity can range from separation of strands within duplex nucleic acids to the physical remodeling or removal of nucleoprotein complexes. In this review, we focus on describing key helicases from the model organism Saccharomyces cerevisiae that contribute to the regulation of homologous recombination, which is an essential DNA repair pathway for fixing damaged chromosomes.

scholarly journals Radiation damage to nucleoprotein complexes in macromolecular crystallography

2015 ◽  
Vol 22 (2) ◽  
pp. 213-224 ◽  
Author(s):  
Charles Bury ◽  
Elspeth F. Garman ◽  
Helen Mary Ginn ◽  
Raimond B. G. Ravelli ◽  
Ian Carmichael ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Radiation Damage ◽  
Diffraction Data ◽  
Dose Range ◽  
Computational Method ◽  
Large Field ◽  
Macromolecular Crystallography ◽  
Damage Mechanisms ◽  
X Ray ◽  
Nucleoprotein Complexes

Significant progress has been made in macromolecular crystallography over recent years in both the understanding and mitigation of X-ray induced radiation damage when collecting diffraction data from crystalline proteins. In contrast, despite the large field that is productively engaged in the study of radiation chemistry of nucleic acids, particularly of DNA, there are currently very few X-ray crystallographic studies on radiation damage mechanisms in nucleic acids. Quantitative comparison of damage to protein and DNA crystals separately is challenging, but many of the issues are circumvented by studying pre-formed biological nucleoprotein complexes where direct comparison of each component can be made under the same controlled conditions. Here a model protein–DNA complex C.Esp1396I is employed to investigate specific damage mechanisms for protein and DNA in a biologically relevant complex over a large dose range (2.07–44.63 MGy). In order to allow a quantitative analysis of radiation damage sites from a complex series of macromolecular diffraction data, a computational method has been developed that is generally applicable to the field. Typical specific damage was observed for both the protein on particular amino acids and for the DNA on, for example, the cleavage of base-sugar N1—C and sugar-phosphate C—O bonds. Strikingly the DNA component was determined to be far more resistant to specific damage than the protein for the investigated dose range. At low doses the protein was observed to be susceptible to radiation damage while the DNA was far more resistant, damage only being observed at significantly higher doses.

scholarly journals Ready separation of proteins from nucleoprotein complexes by reversible modification of lysine residues

1980 ◽  
Vol 191 (1) ◽  
pp. 269-272 ◽  
Author(s):  
J K Shetty ◽  
J E Kinsella
Keyword(s):  
Nucleic Acids ◽  
Nucleoprotein Complex ◽  
Lysine Residues ◽  
Acidic Conditions ◽  
Reversible Modification ◽  
Citraconic Anhydride ◽  
Nucleoprotein Complexes ◽  
Separation Of Proteins ◽  
Phosphate Groups

Modification of proteins with citraconic anhydride altered the electrostatic relationship between cationic epsilon-NH3+ groups of lysine residues of proteins and anionic phosphate groups of nucleic acids, thereby destabilizing the nucleoprotein complex. This procedure facilitated the separation of proteins from nucleic acids at pH4-4.2. The modifying groups were then deacylated from the proteins under acidic conditions (pH3-6) at 30 degrees C.

scholarly journals Srs2 and Pif1 as Model Systems for Understanding Sf1a and Sf1b Helicase Structure and Function

Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1319
Author(s):  
Aviv Meir ◽  
Eric C. Greene
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nucleic Acids ◽  
Model Organism ◽  
Structure And Function ◽  
Model Systems ◽  
Genome Integrity ◽  
Repair Pathway ◽  
Domains Of Life ◽  
Nucleoprotein Complexes ◽  
And Function

Helicases are enzymes that convert the chemical energy stored in ATP into mechanical work, allowing them to move along and manipulate nucleic acids. The helicase superfamily 1 (Sf1) is one of the largest subgroups of helicases and they are required for a range of cellular activities across all domains of life. Sf1 helicases can be further subdivided into two classes called the Sf1a and Sf1b helicases, which move in opposite directions on nucleic acids. The results of this movement can range from the separation of strands within duplex nucleic acids to the physical remodeling or removal of nucleoprotein complexes. Here, we describe the characteristics of the Sf1a helicase Srs2 and the Sf1b helicase Pif1, both from the model organism Saccharomyces cerevisiae, focusing on the roles that they play in homologous recombination, a DNA repair pathway that is necessary for maintaining genome integrity.

SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS IN PROTEIN SOLUTIONS AND IN CRUDE ORGAN EXTRACTS

1955 ◽  
Vol 33 (1) ◽  
pp. 1010-1017
Author(s):  
E. Annau
Keyword(s):  
Absorption Spectrum ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Mouse Liver ◽  
Spectrophotometric Analysis ◽  
Protein Solutions ◽  
Spectrophotometric Procedure ◽  
Acid Protein ◽  
Calf Thymus ◽  
Nucleoprotein Complexes

A spectrophotometric procedure has been presented by which an absorption spectrum, essentially characteristic for nucleic acids, could be obtained from mixed nucleic acid protein solutions, and mouse liver extracts. To examine the efficiency of the method for nucleoprotein complexes, spectra from purified calf thymus nucleohiston were prepared showing the absorption curves of both of its components: nucleic acids and histon.

SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS IN PROTEIN SOLUTIONS AND IN CRUDE ORGAN EXTRACTS

1955 ◽  
Vol 33 (6) ◽  
pp. 1010-1017 ◽  
Author(s):  
E. Annau
Keyword(s):  
Absorption Spectrum ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Mouse Liver ◽  
Spectrophotometric Analysis ◽  
Protein Solutions ◽  
Spectrophotometric Procedure ◽  
Acid Protein ◽  
Calf Thymus ◽  
Nucleoprotein Complexes

A spectrophotometric procedure has been presented by which an absorption spectrum, essentially characteristic for nucleic acids, could be obtained from mixed nucleic acid protein solutions, and mouse liver extracts. To examine the efficiency of the method for nucleoprotein complexes, spectra from purified calf thymus nucleohiston were prepared showing the absorption curves of both of its components: nucleic acids and histon.

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