Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Nicking-closing enzyme assembles nucleosome-like structures in vitro.

J E Germond, J Rouvière-Yaniv, M Yaniv

    Proceedings of the National Academy of Sciences of the United States of America
    |August 1, 1979
    PubMed
    Summary
    This summary is machine-generated.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    V osteotomy and Ilizarov technique for residual idiopathic or neurogenic clubfeet.

    Journal of orthopaedic surgery (Hong Kong)·2008
    Same author

    Characterization of mammary tumors from Brg1 heterozygous mice.

    Oncogene·2007
    Same author

    [Assays on comparing the local concentration of HU protein in the different regions of Escherichia coli genomic DNA].

    Molekuliarnaia biologiia·2005
    Same author

    Congenital absence (ankylosis) of the knee.

    The Journal of bone and joint surgery. British volume·2004
    Same author

    Hepatocyte nuclear factor 1alpha controls the expression of terminal complement genes.

    The Journal of experimental medicine·2001
    Same author

    A yeast homolog of chromatin assembly factor 1 is involved in early ribosome assembly.

    Current biology : CB·2001

    Researchers assembled histone-DNA complexes into nucleosome-like particles, inducing superhelical turns in DNA. This process created beaded structures and DNA fragments characteristic of nucleosomes, mimicking in vivo chromatin organization.

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Structural Biology

    Background:

    • Histones are crucial proteins that package DNA into chromatin.
    • Nucleosomes are the fundamental repeating units of chromatin, essential for genome organization and regulation.
    • Understanding nucleosome assembly is key to comprehending DNA accessibility and gene expression.

    Purpose of the Study:

    • To investigate the in vitro assembly of nucleosome-like particles using core histones and DNA.
    • To determine the structural and topological changes in DNA during nucleosome formation.
    • To characterize the DNA fragments generated from assembled histone-DNA complexes.

    Main Methods:

    • In vitro assembly of histone-DNA complexes using purified core histones (H2A, H2B, H3, H4) and relaxed circular DNA.

    Related Experiment Videos

  • Utilizing a chromatin extract with nicking-closing enzyme activity or the purified enzyme for assembly.
  • Analysis of DNA topology by measuring superhelical turns.
  • Structural characterization using electron microscopy.
  • Enzymatic digestion with micrococcal nuclease to analyze DNA fragment lengths.

    • Main Results:

    • Nucleosome-like particles were successfully assembled at physiological ionic strengths.
    • DNA molecules incorporated into these particles exhibited induced superhelical turns, approaching physiological levels.
    • Electron microscopy revealed beaded structures and reduced DNA contour length in assembled complexes.
    • Micrococcal nuclease digestion produced 145-base-pair fragments, indicative of nucleosome core particles, along with shorter DNA fragments.

    Conclusions:

    • In vitro assembly methods can effectively recapitulate key features of nucleosome formation.
    • The assembly process induces significant topological changes in DNA, essential for chromatin structure.
    • The generated DNA fragments confirm the formation of stable nucleosome core particles and subnucleosomal structures.