The Journal of Immunology, Vol 155, Issue 5 2498-2514, Copyright © 1995 by American Association of Immunologists

ARTICLES

Identification and localization of a developmental stage-specific promoter activity from the murine lambda 5 gene

J Yang, MA Glozak and BB Blomberg
Department of Microbiology and Immunology, University of Miami School of Medicine, FL 33101, USA.

The lambda 5 protein is expressed in pre-B cells in association with VpreB and mu-heavy chains, and is critical for differentiation to the B cell stage. Pre-B cell-specific expression of the lambda 5 and VpreB genes is regulated at the level of transcription initiation. In this report, we have identified several DNase l-hypersensitive sites 2.5- to 6.0-kb downstream of the lambda 5 gene, which are present in the pre-B cell line 70Z/3, but not in the myeloma cell line j558L. These sites, however, were shown to have no transcriptional enhancer activity as measured by transient transfection. Enhancer activity was identified within a 361-bp fragment (-296 to +65, where +1 is the major 5' transcription initiation site) upstream of the mouse lambda 5 gene. This activity is orientation and position independent, and is also tissue and differentiation stage specific (active in pre-B but not B and T cells). Deletion constructs indicate that three adjacent areas (- 210 to -169, -153 to -64, and -64 to -22) are all necessary for enhancer activity. Pre-B cell-specific promoter activity was shown to reside within the -219 to +109 fragment. Basal promoter activity resides within the -64 to +109 fragment, but is not tissue specific or stage specific. A negative element within the -101 to -64 region is active in all lymphoid cell lines tested and therefore cannot by itself be responsible for the tissue and stage specificity. The data indicate that the elements responsible for the enhancer activity (-210 to -22) are part of the lambda 5 gene promoter and likely confer the tissue and stage specificity via positive elements within the -210 to -22 region.

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