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

Nucleoside diphosphokinase activity associated with DNA polymerases.

L K Miller, R D Wells

    Proceedings of the National Academy of Sciences of the United States of America
    |September 1, 1971
    PubMed
    Summary

    Nucleoside diphosphokinase (NDP kinase) activity was found in DNA polymerase preparations from bacteria and viruses. This enzyme activity can lead to the apparent use of diphosphates in DNA synthesis when triphosphates are present.

    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

    Ambulatory care in interventional radiology: a framework for radiology day-case.

    Clinical radiology·2022
    Same author

    The System NaCl-AlCl<sub>3</sub>.

    Journal of research of the National Bureau of Standards. Section A, Physics and chemistry·2020
    Same author

    Long intronic GAA*TTC repeats induce epigenetic changes and reporter gene silencing in a molecular model of Friedreich ataxia.

    Nucleic acids research·2008
    Same author

    Anti- and pro-apoptotic activities of baculovirus and Drosophila IAPs in an insect cell line.

    Cell death and differentiation·2006
    Same author

    Sex difference in the effect of obesity on 24-hour mean serum gonadotropin levels.

    Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme·2003
    Same author

    Chromatin structure of yeast minichromosomes containing triplet repeat sequences associated with human hereditary neurological diseases.

    Nucleic acids research. Supplement (2001)·2003

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Enzymology

    Background:

    • DNA polymerases are crucial enzymes for DNA replication and repair.
    • Nucleoside diphosphokinases (NDP kinases) are enzymes that catalyze the transfer of a phosphate group from nucleoside triphosphates to nucleoside diphosphates.
    • The presence of enzymatic activities other than DNA polymerization in purified DNA polymerase preparations can complicate the interpretation of experimental results.

    Purpose of the Study:

    • To investigate the presence and nature of nucleoside diphosphokinase (NDP kinase) activity in purified DNA polymerase preparations.
    • To determine if NDP kinase activity affects the apparent substrate utilization of DNA polymerases.
    • To characterize the NDP kinase activity associated with DNA polymerase I from Escherichia coli.

    Main Methods:

    Related Experiment Videos

    • Purification of DNA polymerase enzymes from Micrococcus luteus, Escherichia coli, and avian myeloblastosis virus.
    • Enzymatic assays to detect and characterize NDP kinase activity using various ribo- and deoxyribonucleoside di- and triphosphates.
    • Subtilisin cleavage of E. coli DNA polymerase I to identify the fragment responsible for NDP kinase activity.

    Main Results:

    • Highly purified DNA polymerase preparations from M. luteus and E. coli, as well as partially purified avian myeloblastosis virus DNA polymerase, exhibited significant NDP kinase activity.
    • NDP kinase activity was also detected in a 76,000 molecular weight protein fragment of E. coli DNA polymerase I, generated by subtilisin cleavage.
    • The NDP kinase activity in M. luteus DNA polymerase preparations utilized a broad range of ribonucleoside and deoxyribonucleoside di- and triphosphates as substrates.
    • The presence of NDP kinase activity led to the apparent incorporation of deoxyribonucleoside diphosphates into DNA, provided a triphosphate was available as a phosphate donor.

    Conclusions:

    • DNA polymerase preparations can contain associated NDP kinase activity.
    • This associated NDP kinase activity can influence the interpretation of DNA synthesis assays by causing the apparent utilization of diphosphates.
    • The findings highlight the importance of assessing enzyme purity and potential contaminating activities in biochemical studies of DNA polymerases.