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

DNA-dependent RNA polymerase from Halobacterium halobium

W Zillig, K O Stetter, M Tobien

    European Journal of Biochemistry
    |November 2, 1978
    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

    Isolation, taxonomy and phylogeny of hyperthermophilic microorganisms.

    World journal of microbiology & biotechnology·2014
    Same author

    Pyrodictium gen. nov., a New Genus of Submarine Disc-Shaped Sulphur Reducing Archaebacteria Growing Optimally at 105°C.

    Systematic and applied microbiology·2012
    Same author

    The Archaebacterium Thermofilum pendens Represents, a Novel Genus of the Thermophilic, Anaerobic Sulfur Respiring Thermoproteales.

    Systematic and applied microbiology·2012
    Same author

    The Archaebacterium Thermococcus celer Represents, a Novel Genus within the Thermophilic Branch of the Archaebacteria.

    Systematic and applied microbiology·2012
    Same author

    Posttranscriptional modification of transfer RNA in the submarine hyperthermophile Pyrolobus fumarii.

    Nucleic acids symposium series·2003
    Same author

    Remarkable morphological diversity of viruses and virus-like particles in hot terrestrial environments.

    Archives of virology·2002

    Researchers purified the DNA-dependent RNA polymerase core enzyme from Halobacterium halobium. This enzyme is crucial for RNA synthesis and its fidelity is confirmed by specific nucleotide requirements.

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Microbiology

    Background:

    • DNA-dependent RNA polymerase is essential for gene expression in all domains of life.
    • Halobacterium halobium, an archaeon, possesses unique molecular mechanisms for transcription.

    Purpose of the Study:

    • To isolate and characterize the DNA-dependent RNA polymerase core enzyme from Halobacterium halobium.
    • To investigate the subunit composition and catalytic properties of the archaeal RNA polymerase.

    Main Methods:

    • Enzyme purification using a multi-step approach including glycerol stabilization, DEAE-cellulose chromatography, streptomycin sulfate precipitation, Biogel chromatography, and heparin affinity chromatography.
    • Analysis of subunit composition via molecular weight estimation.
    • RNA synthesis assays using native and denatured DNA templates, with and without a transcription factor.

    Related Experiment Videos

    Main Results:

    • Successfully isolated a stable DNA-dependent RNA polymerase core enzyme from Halobacterium halobium.
    • The enzyme comprises four or five subunits, with estimated molecular weights and potential heterogeneity in the 49,000-Mr subunit.
    • Denatured DNA served as a more efficient template than native DNA for RNA synthesis.
    • A sigma-like transcription factor was identified that specifically stimulates the transcription of native DNA.

    Conclusions:

    • The purified Halobacterium halobium RNA polymerase core enzyme is functional and requires a DNA template for RNA synthesis.
    • The enzyme's subunit composition and the identification of a transcription factor provide insights into archaeal transcription regulation.
    • The fidelity of transcription was demonstrated by the requirement for UTP alongside ATP with a poly[d(A-T)] template.