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

Ribonuclease H: from discovery to 3D structure.

R J Crouch1

  • 1Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.

The New Biologist
|September 1, 1990
PubMed
Summary

Ribonucleases H (RNases H) from E. coli and retroviruses share conserved active sites for RNA hydrolysis. These enzymes, crucial for replication and DNA synthesis, may function as part of larger proteins.

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

Actin and myosin contribute to mammalian mitochondrial DNA maintenance.

Nucleic acids research·2011
Same author

RNase H1 of Saccharomyces cerevisiae: methods and nomenclature.

Methods in enzymology·2001
Same author

Quantitative regulation of class switch recombination by switch region transcription.

The Journal of experimental medicine·2001
Same author

HIV-1 reverse transcriptase interaction with model RNA-DNA duplexes.

Analytical biochemistry·2001
Same author

Sorting of mannose 6-phosphate receptors mediated by the GGAs.

Science (New York, N.Y.)·2001
Same author

Signal-binding specificity of the mu4 subunit of the adaptor protein complex AP-4.

The Journal of biological chemistry·2001

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Ribonucleases H (RNases H) are enzymes with conserved features in both prokaryotic (Escherichia coli) and retroviral systems.
  • RNase H plays critical roles in DNA replication initiation in E. coli and in retroviral DNA synthesis.
  • Multiple forms of RNase H exist across prokaryotes and eukaryotes, suggesting diverse biological functions.

Purpose of the Study:

  • To investigate the conserved structural and functional characteristics of Ribonucleases H.
  • To elucidate the active site of E. coli RNase H through crystallographic studies.
  • To explore the potential for RNase H to be integrated into larger polypeptide complexes.

Main Methods:

  • Comparative analysis of primary amino acid sequences of RNases H.
  • Activity assays to determine enzymatic function.
  • X-ray crystallography of E. coli RNase H to resolve its three-dimensional structure.

Main Results:

  • Identified shared primary amino acid sequence and activity features between E. coli and retroviral RNases H.
  • Crystallographic data revealed that conserved amino acids form the active site for RNA hydrolysis in RNA-DNA hybrids.
  • Evidence suggests RNase H can be part of larger proteins, influencing its localization and activity.

Conclusions:

  • The active site of RNase H is highly conserved, indicating a fundamental role in nucleic acid metabolism.
  • The presence of multiple RNase H forms suggests specialized roles in different cellular contexts.
  • RNase H activity can be modulated by its integration into larger protein complexes, as seen in retroviral reverse transcriptase.

Related Experiment Videos