Journal
MOLECULAR AND CELLULAR BIOLOGY
Volume 25, Issue 8, Pages 3220-3231Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/MCB.25.8.3220-3231.2005
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Funding
- NIGMS NIH HHS [R01 GM029935, GM59809, R01 GM059809, GM29935] Funding Source: Medline
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The mouse immunoglobulin kappa (Ig kappa) gene contains an intronic enhancer and two enhancers downstream of its transcription unit. Using chromosome conformation capture technology, we demonstrate that rearranged and actively transcribed Ig kappa alleles in MPC-11 plasmacytoma cells exhibit mutual interactions over 22 kb between these three enhancers and V kappa gene promoters. In addition, the 5' region of the active transcription unit exhibits a continuum of interactions with downstream chromatin segments. We also observe interactions between Ei and EX with 3' boundary sequences 24 kb downstream of Ed, adjacent to a neighboring housekeeping gene. Very similar interactions between the enhancers are also exhibited by normal B cells isolated from mouse splenic tissue but not by germ line transcriptionally inactive alleles of T cells or P815 mastocytoma cells, which exhibit a seemingly linear chromatin organization. These results fit a looping mechanism for enhancer function like in the P-globin locus and suggest a dynamic modulation of the spatial organization of the active Ig kappa locus. Chromatin immunoprecipitation experiments reveal that the interacting Ig kappa gene cis-acting sequences are associated with AP-4, E47, and p65NF-kappa B, potential protein candidates that may be responsible for initiating and/or maintaining the formation of these higher-order complexes. However, S107 plasmacytoma cells that lack NF-kappa B still exhibit mutual interactions between the Ig kappa gene enhancers.
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