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Related Experiment Videos

Transcription-dependent R-loop formation at mammalian class switch sequences.

R B Tracy1, M R Lieber

  • 1Department of Pathology, Norris Comprehensive Cancer Center, University of Southern California Keck School of Medicine, 1441 Eastlake Avenue, Los Angeles, CA 90089-9176, USA.

The EMBO Journal
|March 4, 2000
PubMed
Summary
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Immunoglobulin class switching involves RNA-DNA hybrids at switch sequences. These structures form R-loops, with a G-rich RNA strand binding the C-rich DNA, facilitating recombination.

Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • Immunoglobulin class switching is a key adaptive immune process.
  • This process relies on recombination at specific switch (S) regions.
  • Transcription through S regions is linked to recombination targeting.

Purpose of the Study:

  • To define the in vitro structure of RNA-DNA hybrids at mouse switch sequences.
  • To elucidate the role of these structures in immunoglobulin class switching.

Main Methods:

  • In vitro analysis of switch sequences (Smicro, Sgamma3, Sgamma2b).
  • DNA/RNA hybrid structure determination using probe sensitivity assays.

Main Results:

  • Identified R-loop formation at mouse switch sequences.

Related Experiment Videos

  • Demonstrated G-rich RNA strand forms a heteroduplex with the C-rich DNA strand.
  • Showed the G-rich DNA strand is primarily single-stranded.

    • Conclusions:

    • The R-loop structure is precisely organized at switch sequences.
    • This organized R-loop is proposed to be essential for recruiting the class switch recombination machinery.