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

Template restriction in human chromatin.

W R Crain, G F Saunders

    Cell Differentiation
    |March 11, 1976
    PubMed
    Summary
    This summary is machine-generated.

    RNA polymerase transcribes human DNA and chromatin. Chromatin transcripts are shorter and initiate differently, suggesting DNA accessibility impacts transcription in human cells.

    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

    Activation of the human PAX6 gene through the exon 1 enhancer by transcription factors SEF and Sp1.

    Nucleic acids research·2001
    Same author

    Missense mutation at the C-terminus of PAX6 negatively modulates homeodomain function.

    Human molecular genetics·2001
    Same author

    Transcriptional regulation of the androgen signaling pathway by the Wilms' tumor suppressor gene WT1.

    Anticancer research·2001
    Same author

    The human sex-determining gene SRY is a direct target of WT1.

    The Journal of biological chemistry·2001
    Same author

    Modulation of PAX6 homeodomain function by the paired domain.

    The Journal of biological chemistry·2000
    Same author

    Mutation in the PAX6 gene in twenty patients with aniridia.

    Human mutation·2000
    Same journal

    Presence of serotonin in early chick embryos.

    Cell differentiation·1988
    Same journal

    Differential expression of the laminin A and B chains in chimeric kidneys.

    Cell differentiation·1988
    Same journal

    Glomerular sialoconjugates of developing and mature rat kidneys.

    Cell differentiation·1988
    Same journal

    Stress proteins by zinc ions in sea urchin embryos.

    Cell differentiation·1988
    Same journal

    Cell differentiation as assayed by the topography and number of ribosomal genes.

    Cell differentiation·1988
    Same journal

    A monoclonal antibody recognizing a common antigen on neurons and fibroblasts in chicken and quail.

    Cell differentiation·1988
    See all related articles

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • DNA-dependent RNA polymerase activity is crucial for gene expression.
    • Chromatin structure can regulate DNA accessibility and transcription.
    • Understanding transcription from different DNA templates is key to gene regulation studies.

    Purpose of the Study:

    • To compare RNA synthesis from purified human DNA versus human leukocyte chromatin.
    • To investigate differences in RNA chain length, initiation sites, and hybridization kinetics.
    • To elucidate the impact of chromatin structure on transcription by E. coli RNA polymerase.

    Main Methods:

    • In vitro transcription using E. coli DNA-dependent RNA polymerase.
    • Labeling RNA 5' termini with 32P-ATP/GTP and internally with 3H-UTP.

    Related Experiment Videos

  • Quantifying RNA chain length via nucleotide incorporation ratios.
  • Hybridization kinetics of RNA transcripts to DNA, assessing RNase resistance.

    • Main Results:

    • Transcripts from chromatin were significantly shorter (2.5x) than those from purified DNA.
    • The ratio of ATP to GTP initiation varied, being higher for chromatin templates.
    • RNA from purified DNA primarily hybridized to nonrepetitive sequences.
    • Chromatin transcripts showed increased RNase resistance of 5' termini at low Cot values during hybridization.

    Conclusions:

    • Chromatin structure restricts RNA polymerase access, leading to shorter transcripts.
    • Template accessibility influences RNA initiation site selection.
    • Chromatin-bound DNA may involve regulatory mechanisms affecting transcription initiation and early elongation.