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

Human replication protein A binds single-stranded DNA in two distinct complexes

L J Blackwell1, J A Borowiec

  • 1Department of Biochemistry, New York University Medical Center, New York 10016.

Molecular and Cellular Biology
|June 1, 1994
PubMed
Summary

Human replication protein A (hRPA) binds single-stranded DNA (ssDNA) in two distinct modes: a cooperative 8-10 nucleotide complex and a 30-nucleotide complex, suggesting flexible roles in DNA metabolism.

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

Nucleolin inhibits Hdm2 by multiple pathways leading to p53 stabilization.

Oncogene·2006
Same author

Distinct MutS DNA-binding modes that are differentially modulated by ATP binding and hydrolysis.

The Journal of biological chemistry·2001
Same author

DNA chain length dependence of formation and dynamics of hMutSalpha.hMutLalpha.heteroduplex complexes.

The Journal of biological chemistry·2001
Same author

5' --> 3' molecular polarity of human replication protein A (hRPA) binding to pseudo-origin DNA substrates.

Biochemistry·2000
Same author

Formation of a complex between nucleolin and replication protein A after cell stress prevents initiation of DNA replication.

The Journal of cell biology·2000
Same author

Replication protein A (RPA): the eukaryotic SSB.

Critical reviews in biochemistry and molecular biology·1999

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Human replication protein A (hRPA) is crucial for DNA replication, repair, and recombination.
  • hRPA interacts with proteins regulating replication, transcription, and cell growth.
  • Understanding hRPA's ssDNA binding is key to its diverse cellular functions.

Purpose of the Study:

  • To investigate the binding characteristics of hRPA to single-stranded DNA (ssDNA).
  • To elucidate the different modes of hRPA-ssDNA complex formation.
  • To explore how these binding modes influence hRPA's role in DNA metabolism.

Main Methods:

  • Gel shift assays were used to analyze hRPA binding to ssDNA of varying lengths.
  • Glutaraldehyde cross-linking detected distinct multimeric hRPA complexes.

Related Experiment Videos

  • Titration of ssDNA length determined minimum binding site sizes.

    • Main Results:

    • hRPA forms at least two distinct complexes with ssDNA: hRPA8-10nt and hRPA30nt.
    • The hRPA8-10nt complex involves cooperative binding with a minimum site size of 8-10 nucleotides.
    • The hRPA30nt complex binds ssDNA in approximately 30-nucleotide steps, coexisting with the smaller complex.

    Conclusions:

    • hRPA exhibits dual ssDNA binding modes, suggesting adaptability in its functions.
    • The ability to form distinct complexes may modulate hRPA's roles in DNA replication, repair, and recombination.
    • These findings provide insight into the regulatory mechanisms of hRPA in DNA metabolism.