Macromolecular crowding allows blunt-end ligation by DNA ligases from rat liver or Escherichia coli.

@article{Zimmerman1983MacromolecularCA,
  title={Macromolecular crowding allows blunt-end ligation by DNA ligases from rat liver or Escherichia coli.},
  author={Steven B. Zimmerman and Barbara H. Pheiffer},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={1983},
  volume={80 19},
  pages={
          5852-6
        }
}
  • S. Zimmerman, B. H. Pheiffer
  • Published 1983
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
In the presence of high concentrations of any of several types of macromolecules, DNA ligase preparations from rat liver nuclei or from Escherichia coli actively catalyze the blunt-end ligation of DNA. This is in contrast to the lack of activity on such substrates by these enzymes under conventional assay conditions. In addition, the previously established activity of T4 DNA ligase on blunt-ended molecules is greatly increased in the presence of high concentrations of macromolecules. Because… Expand

Figures and Topics from this paper

Blunt-end and single-strand ligations by Escherichia coli ligase: influence on an in vitro amplification scheme.
TLDR
The results of this study indicate that E. coli ligase also joins blunt-ended DNA molecules and some single-stranded oligodeoxyribonucleotides, in the absence of a complementary template, with an efficiency which is sensitive to both the concentrations of DNA substrate and enzyme. Expand
DNA Ligases
  • Stanley Tabor
  • Chemistry, Medicine
  • Current protocols in molecular biology
  • 2001
TLDR
Two ligases are used for nucleic acid research and their reaction conditions and applications are described in this unit; under normal reaction conditions, only T4 DNA ligase will ligate blunt ends. Expand
Template‐independent ligation of single‐stranded DNA by T4 DNA ligase
TLDR
Comparison of ligation yields obtained with substrates differing in their strandedness at the terminal segments involved in ligation shows that an acceptor duplex DNA segment bearing a 3′‐hydroxy end, but lacking a 5′‐phosphate end, is sufficient to play a role as a cofactor in blunt‐end ligation. Expand
Specificity of the nick-closing activity of bacteriophage T4 DNA ligase.
TLDR
These properties of T4 DNA ligase for oligo pairs complementary to the beta-globin gene at the sequence surrounding the single bp mutation responsible for sickle-cell anemia are demonstrated. Expand
Analysis of ligation and DNA binding by Escherichia coli DNA ligase (LigA).
TLDR
The over-expression and purification of various fragments of E. coli LigA allowed the investigation of the different domains in DNA-binding and ligation by this enzyme, finding the BRCT domain can bind DNA, but it is not essential for DNA nick-joining activity in vitro or in vivo. Expand
DNA ligases from rat liver. Purification and partial characterization of two molecular forms.
TLDR
Rat liver DNA ligase II has a lower Km for ATP than other eukaryotic DNA ligases, and can use ATP alpha S as a cofactor in the ligation reaction much more efficiently thanDNA ligase I, further discriminating the ATP binding sites of these enzymes. Expand
Ultrafast spectroscopy on DNA-cleavage by endonuclease in molecular crowding.
TLDR
The enhanced dynamical flexibility of protein in presence of PEG as revealed from excited state lifetime and polarization gated anisotropy of ANS has been correlated with the stronger DNA-binding for the higher nuclease activity. Expand
Regulation of DNA nucleases by molecular crowding
TLDR
The hydrolysis of a 29-mer double-stranded DNA by the endonucleases DNase I and S1 nuclease was substantially enhanced by molecular crowding using polyethylene glycol (PEG); however, molecularrowding had little effect on Hydrolysis by exo III and exo I exonuclease. Expand
Effect of molecular crowding on DNA polymerase activity
TLDR
Analysis of the effect of molecular crowding on DNA polymerase activity using polyethylene glycol PEG of various molecular weights as a crowding agent suggests that quantitative measurements of biomolecular structure and function are useful for understanding the Behavior in vivo and for biotechnology applications in vitro. Expand
Effect of histone H1, poly(ethyleneglycol) and DNA concentration on intermolecular and intramolecular ligation by T4 DNA ligase.
TLDR
It is shown that histone H1, which is involved in the formation of chromatin fibers, is able to stimulate intermolecular ligation by T4 ligase, and it is suggested that salts enhance the hydrophobic interactions between ligase and DNA that take place in the presence of poly(ethyleneglycol). Expand
...
1
2
3
4
5
...

References

SHOWING 1-3 OF 3 REFERENCES
Patterns of integration of DNA microinjected into cultured mammalian cells: evidence for homologous recombination between injected plasmid DNA molecules.
TLDR
It was found that even linear plasmid DNA was recruited into the concatemer by homologous recombination, even though cultured mammalian cells contain the enzymes for ligating two DNA molecules very efficiently irrespective of the sequences or topology at their ends. Expand
Molecular Cloning: A Laboratory Manual
TLDR
The content has been entirely recast to include nucleic-acid based methods selected as the most widely used and valuable in molecular and cellular biology laboratories. Expand
Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
  • J. Sutcliffe
  • Biology, Medicine
  • Cold Spring Harbor symposia on quantitative biology
  • 1979