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

A single mutation, RecB(D1080A,) eliminates RecA protein loading but not Chi recognition by RecBCD enzyme.

D G Anderson1, J J Churchill, S C Kowalczykowski

  • 1Sections of Microbiology and of Molecular and Cellular Biology, University of California, Davis, California 95616-8665, USA.

The Journal of Biological Chemistry
|September 10, 1999
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

Immune-remodeling mRNAs expressing IRF8 or NIK generate durable antitumor immunity in multiple cancer models.

Nature biotechnology·2026
Same author

Augmentation of Pedicle Screws Using Bone Grafting in Patients with Spinal Osteoporosis.

Sovremennye tekhnologii v meditsine·2022
Same author

Intraosseous basivertebral nerve ablation for the treatment of chronic low back pain: a prospective randomized double-blind sham-controlled multi-center study.

European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society·2018
Same author

South African guideline on deep brain stimulation for Parkinson's disease.

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde·2017
Same author

ALCOHOL DEHYDROGENASE AND GLYCEROL-3-PHOSPHATE DEHYDROGENASE CLINES IN DROSOPHILA MELANOGASTER ON DIFFERENT CONTINENTS.

Evolution; international journal of organic evolution·2017
Same author

Poly(glycoamidoamine) brush nanomaterials for systemic siRNA delivery in vivo.

Biomaterials science·2016
Same journal

YhbO is a DJ-1 family glyoxalase and α-oxoaldehyde hydratase that confers resistance to reactive carbonyl stress (112).

The Journal of biological chemistry·2026
Same journal

ARMH3 acts as a central scaffold at the Golgi/TGN through interactions with Arl5, GBF1, and PI4KB.

The Journal of biological chemistry·2026
Same journal

PAX8 controls proximal tubule epithelial identity and stress response through epigenetic modification of distal regulatory elements.

The Journal of biological chemistry·2026
Same journal

Saturated cardiolipins are potent disruptors of inner mitochondrial membrane structure and function.

The Journal of biological chemistry·2026
Same journal

Phosphate release from myosin Va occurs after the initial powerstroke but before the secondary powerstroke associated with ADP-release.

The Journal of biological chemistry·2026
Same journal

Epigenetic silencing of miR-141 via core promoter methylation is associated with short-term bladder cancer progression.

The Journal of biological chemistry·2026
See all related articles

The RecBCD enzyme initiates DNA repair in E. coli. A specific mutation prevents RecBCD from loading RecA protein, impacting DNA repair, but it still responds to Chi sites.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Homologous recombination and double-stranded DNA break repair in Escherichia coli are initiated by the RecBCD enzyme.
  • RecBCD unwinds and degrades DNA, with processing regulated by the Chi recombination hot spot.
  • Chi induces a polarity switch in DNA degradation and activates RecBCD for RecA loading.

Purpose of the Study:

  • To investigate the function of a RecBCD enzyme mutant (RecB(D1080A)CD) lacking nuclease activity.
  • To determine the role of the RecB(D1080A)CD mutation in RecA protein loading and Chi site response.
  • To identify the specific locus within RecBCD essential for nuclease function and RecA loading coordination.

Main Methods:

  • Biochemical assays to assess helicase and nuclease activity of the RecB(D1080A)CD enzyme.

Related Experiment Videos

  • In vitro studies to evaluate RecA protein loading onto DNA substrates.
  • Analysis of RecB(D1080A)CD enzyme inactivation in the presence and absence of Chi sites.

    • Main Results:

    • The RecB(D1080A)CD enzyme functions as a processive helicase but lacks nuclease activity.
    • RecB(D1080A)CD is unable to coordinate RecA protein loading, irrespective of Chi site presence.
    • RecB(D1080A)CD exhibits Chi-dependent inactivation, indicating a response to Chi sites.

    Conclusions:

    • A specific locus in RecBCD is crucial for both nuclease activity and coordinating RecA loading.
    • The RecB(D1080A)CD mutation disrupts RecA loading coordination while retaining Chi site responsiveness.
    • These findings elucidate the distinct roles of different RecBCD enzyme domains in DNA repair pathways.