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 replication in vitro by recombinant DNA-polymerase-alpha-primase

F Stadlbauer1, A Brueckner, C Rehfuess

  • 1Institut für Biochemie d. LMU München, Germany.

European Journal of Biochemistry
|June 15, 1994
PubMed
Summary

Researchers characterized the DNA-polymerase-alpha--primase complex, identifying subunits responsible for primase and polymerase activities. This study elucidates the complex

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

Sequence analysis of proteolytic fragments of 124-kilodalton phytochrome from etiolatedAvena sativa L.: Conclusions on the conformation of the native protein.

Planta·2013
Same author

White matter integrity in physically fit older adults.

NeuroImage·2013
Same author

Sorafenib tolerability in elderly patients with advanced renal cell carcinoma: results from a large pooled analysis.

British journal of cancer·2013
Same author

For knowledge of the so-called afterimage.

Schweizerische medizinische Wochenschrift·2010
Same author

Pooled safety analysis of BAY 43-9006 (sorafenib) monotherapy in patients with advanced solid tumours: Is rash associated with treatment outcome?

European journal of cancer (Oxford, England : 1990)·2006
Same author

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells.

Leukemia·2005

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Enzymology

Background:

  • The DNA-polymerase-alpha--primase complex is essential for DNA replication.
  • This complex comprises four subunits: p180, p68, p58, and p48.
  • It possesses both DNA primase and DNA polymerase activities.

Purpose of the Study:

  • To clone and characterize the subunits of the human and mouse DNA-polymerase-alpha--primase complex.
  • To investigate the biochemical properties of individual subunits and their complexes.
  • To determine the enzymatic activities of recombinant DNA-polymerase-alpha--primase complexes.

Main Methods:

  • Cloning of cDNAs encoding DNA-polymerase-alpha--primase subunits from human and mouse.
  • Over-expression of single polypeptides and co-expression of subunit complexes using baculovirus vectors.

Related Experiment Videos

  • Purification of proteins and biochemical characterization of their enzymatic activities.

    • Main Results:

    • High amino acid conservation was observed among mammalian proteins.
    • The mouse p48 subunit (Mp48) alone exhibited primase activity.
    • Co-expression of Mp48 and Mp58 subunits yielded a stable, highly active DNA primase.
    • Co-expression of all four subunits produced substantial amounts of tetrameric DNA-polymerase-alpha--primase.
    • Trimeric and tetrameric complexes demonstrated both primase and polymerase activities.
    • Tetrameric recombinant DNA-polymerase-alpha--primase efficiently synthesized double-stranded M13 DNA and replicated polyoma viral DNA in vitro.

    Conclusions:

    • The p48 subunit is crucial for primase activity.
    • The DNA-polymerase-alpha--primase complex can be reconstituted from its subunits.
    • Recombinant DNA-polymerase-alpha--primase exhibits robust enzymatic functions essential for DNA replication.