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

Mutational analysis on structure-function relationship of a holliday junction specific endonuclease RuvC

K Ichiyanagi1, H Iwasaki, T Hishida

  • 1Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
|November 13, 1998
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

Supernumerary ring chromosomes and nuclear blebs in some low-grade malignant soft tissue tumours: atypical lipomatous tumours and dermatofibrosarcoma protuberans.

Virchows Archiv : an international journal of pathology·1998
Same author

Abortive recombination in Escherichia coli ruv mutants blocks chromosome partitioning.

Genes to cells : devoted to molecular & cellular mechanisms·1998
Same author

Reirradiation therapy for brain metastases from small cell lung cancer.

Radiation medicine·1998
Same author

Remission induction by granulocyte colony-stimulating factor in hypoplastic acute myelogenous leukemia complicated by infection. A case report and review of the literature.

Acta haematologica·1998
Same author

[The efficacy of low-dose propofol for intrathecal morphine-induced pruritus].

Masui. The Japanese journal of anesthesiology·1998
Same author

[Preventive effect of fluid warmer system on hypothermia induced by rapid intravenous infusion].

Masui. The Japanese journal of anesthesiology·1998
Same journal

The Nuclear Export Signal of IκBα Drives RelB Oscillations in the Noncanonical NF-κB Pathway.

Genes to cells : devoted to molecular & cellular mechanisms·2026
Same journal

The SQSTM1 L341V Variant Associated With Sporadic ALS Promotes the Accumulation of Enlarged Ubiquitin-Positive SQSTM1 Bodies.

Genes to cells : devoted to molecular & cellular mechanisms·2026
Same journal

Partial Depletion of SART1 Selectively Induces Mitotic Defects and Cell Death in Cancer Cells.

Genes to cells : devoted to molecular & cellular mechanisms·2026
Same journal

FOXO1 Is Required for Growth and Viability of Cancer-Associated Fibroblasts in Human Breast Carcinomas.

Genes to cells : devoted to molecular & cellular mechanisms·2026
Same journal

Strain-Dependent Functional Variation of a Sex Specific Sox9 Enhancer.

Genes to cells : devoted to molecular & cellular mechanisms·2026
Same journal

An Outflow Tract Myocardium-Specific Enhancer at the Sema3c Locus During Heart Development.

Genes to cells : devoted to molecular & cellular mechanisms·2026
See all related articles

Random mutagenesis identified key residues in Escherichia coli RuvC protein, essential for Holliday junction resolution during homologous recombination. Dimer formation and specific interactions are critical for RuvC function.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Escherichia coli RuvC protein is a crucial endonuclease for resolving Holliday junctions in homologous recombination.
  • RuvC function involves dimerization, DNA binding, and endonucleolytic cleavage, with its crystal structure previously determined.

Purpose of the Study:

  • To identify functionally critical residues of RuvC through random mutagenesis.
  • To elucidate the roles of specific residues in RuvC's catalytic activity, dimerization, and DNA binding.

Main Methods:

  • Random mutagenesis of ruvC genes.
  • In vivo and in vitro characterization of mutant RuvC proteins.
  • Analysis of dimerization, DNA binding, and cleavage activities.

Related Experiment Videos

Main Results:

  • Mutations frequently occurred at the four acidic residues of the catalytic center.
  • Several mutants affected the dimer interface; one failed to dimerize, while others dimerized but were cleavage-defective.
  • F69L and K118R mutants could not cleave junctions despite dimer formation and DNA binding.

Conclusions:

  • Random mutagenesis revealed structurally and functionally important, conserved RuvC residues.
  • Dimer formation and intact subunit interactions are vital for RuvC's junction binding and cleavage.
  • Phenylalanine-69 and Lysine-118 are critical for junction cleavage activity.