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

Mapping pseudouridines in RNA molecules.

J Ofengand1, M Del Campo, Y Kaya

  • 1Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Miami, Florida 33101, USA. jofengan@molbio.med.miami.edu

Methods (San Diego, Calif.)
|February 28, 2002
PubMed
Summary

Pseudouridine, a modified nucleotide, enhances RNA structure and interactions. A new method accurately maps pseudouridine locations in various RNA molecules, aiding in understanding its function.

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

Gastrointestinal manifestations in patients with gastric adenocarcinoma and proximal polyposis of the stomach (GAPPS): a systematic review with analysis of individual patient data.

Hereditary cancer in clinical practice·2024
Same author

Molecular profiling of bacterial blight resistance in Malaysian rice cultivars.

Brazilian journal of biology = Revista brasleira de biologia·2022
Same author

Plasma proteome profiling identifies changes associated to AD but not to FTD.

Acta neuropathologica communications·2022
Same author

RELATION OF APELIN, TUMOR NECROSIS FACTOR ALPHA AND CLAUDIN-5 TO BODY MASS INDEX IN CHOLECYSTECTOMIES.

Acta endocrinologica (Bucharest, Romania : 2005)·2022
Same author

Trajectories of prenatal alcohol exposure and behavioral outcomes: Findings from a community-based sample.

Drug and alcohol dependence·2022
Same author

Seizures and electroencephalography findings in 61 patients with fetal alcohol spectrum disorders.

European journal of medical genetics·2016

Area of Science:

  • Biochemistry
  • Molecular Biology
  • RNA Biology

Background:

  • Pseudouridine is a modified nucleotide found in various structured RNAs, including ribosomal RNA (rRNA), transfer RNA (tRNA), and small nuclear RNAs.
  • Its formation involves enzyme-catalyzed isomerization of uridine residues post-transcription, creating an additional hydrogen bond donor at C-5.
  • This modification is hypothesized to stabilize RNA conformations and RNA-RNA interactions.

Purpose of the Study:

  • To develop and describe a method for mapping the location and number of pseudouridine modifications in RNA molecules.
  • To enable a broader survey of pseudouridine distribution across different organisms and RNA types.

Main Methods:

  • The method utilizes carbodiimide adduct formation with uridine (U), guanosine (G), and pseudouridine.

Related Experiment Videos

  • Selective alkali treatment removes adducts from U and G but not from pseudouridine's N-3 position.
  • This differential modification leads to attenuated primed reverse transcription, generating a stop band one residue 3' to pseudouridine on sequencing gels.

    • Main Results:

    • The described method simplifies the mapping of pseudouridines in both large RNAs (like rRNA) and small RNAs (like tRNA).
    • The technique does not require purified RNAs, only knowledge of the primary sequence, and can scan approximately 30-40 residues from the 3' end.
    • A poly(A) tailing procedure extends the mapping capability to within a few nucleotides of the 3' terminus.

    Conclusions:

    • This novel mapping approach significantly facilitates the study of pseudouridine distribution and abundance in RNA.
    • Understanding pseudouridine's location is crucial for elucidating its functional roles in RNA structure and interactions.
    • The method's adaptability allows for comprehensive analysis across diverse RNA species and organisms.