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Related Experiment Videos

Recombination potential of the human DIR elements

B B Moore1, K Meek

  • 1Harold C. Simmons Arthritis Research Center, University of Texas Southwestern Medical Center, Dallas 75235.

Journal of Immunology (Baltimore, Md. : 1950)
|March 1, 1995
PubMed
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Human DIR genes, involved in immunoglobulin gene rearrangement, show minimal participation in DH-DH and DH-JH rearrangements. PCR analysis confirms that conventional DH-DH rearrangements are more prevalent than those involving DIR elements.

Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • Human DIR genes are GC-rich elements situated between DH gene segments (DN and DM).
  • DIR genes possess recombination signal sequences (RSS) potentially enabling rearrangements that adhere to the 12/23 recombination rule.
  • Existing assignments of immunoglobulin CDR3 sequences to DIR usage exhibit inconsistencies, suggesting alternative explanations like N segment addition.

Purpose of the Study:

  • To investigate the recombination potential of human DIR elements.
  • To analyze the frequency and mechanisms of DIR gene involvement in immunoglobulin heavy chain gene rearrangement.
  • To clarify the role of DIR-associated RSS in DH-DH and DH-JH rearrangements.

Main Methods:

  • Polymerase Chain Reaction (PCR) was employed to analyze the recombination potential of human DIR elements.

Related Experiment Videos

  • Amplification strategies were designed to detect both DH-DIR and DH-DH rearrangements, as well as DH-JH rearrangements.
  • Analysis focused on rearrangements involving DIR elements and flanking DH genes (DM and DN).

    • Main Results:

    • DH-JH rearrangements of flanking DH genes (DM and DN) were readily detected.
    • Very limited evidence for DIR-JH rearrangement was found.
    • In PCR assays designed to detect both DH-DIR and DH-DH rearrangements, DH-DH rearrangements predominated, indicating a preference for rearrangements that violate the 12/23 rule over those involving DIR elements.

    Conclusions:

    • The DIR-associated RSS appear to play a minimal role in both DH-DH and DH-JH rearrangements.
    • Conventional DH-DH rearrangements occur frequently during human immunoglobulin gene rearrangement.
    • The findings challenge the proposed role of DIR elements in generating Ig CDR3 diversity via standard recombination pathways.