Immunoglobulin gene expression requires rearrangement of heavy and light chain gene segments during B cell development, that brings promoter elements into the proximity of enhancer elements. The focus of this proposal is to continue to study events that control Ig gene rearrangement and expression early in B cell ontogeny, in both mouse and human. This includes characterization of elements which regulate germline transcription of the kappa locus, and their potential role in targeting gene segments for rearrangement. The developmental activation of the kappa intron and 3' enhancers will be examined by transgenic mouse studies. The specific role of the intron enhancer will be determined by examining the effects of enhancer deletion in A-MuLV transformed cell lines that normally rearrange kappa genes, and ES cells used to generate genetically altered mice. The structural and functional components that comprise the kappa intron enhancer will be studied by defining sequences and protein-DNA interactions that contribute to transcriptional activation and suppression in model cell culture systems. This includes cloning a novel, inducible factor that binds a region in the enhancer, as well as defining requirements for spatial arrangements of all the sequence motifs that contribute to full enhancer activity. Sequences that flank the core intron enhancer and act as silencers of enhancer activity will be characterized. A number of these studies will involve direct comparisons of mouse and human enhancer organization and function. Finally, an ongoing collaborative study will be maintained to study the T cell dependent, heavy chain enhancer-mediated suppression of IgE expression in murine hybridomas.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM037687-12
Application #
2178895
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1987-09-15
Project End
1995-11-30
Budget Start
1993-12-01
Budget End
1995-11-30
Support Year
12
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Biochemistry
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
O'Brien, D P; Oltz, E M; Van Ness, B G (1997) Coordinate transcription and V(D)J recombination of the kappa immunoglobulin light-chain locus: NF-kappaB-dependent and -independent pathways of activation. Mol Cell Biol 17:3477-87
Prabhu, A; O'Brien, D P; Weisner, G L et al. (1996) Octamer independent activation of transcription from the kappa immunoglobulin germline promoter. Nucleic Acids Res 24:4805-11
Schanke, J T; Marcuzzi, A; Podzorski, R P et al. (1994) An AP1 binding site upstream of the kappa immunoglobulin intron enhancer binds inducible factors and contributes to expression. Nucleic Acids Res 22:5425-32
Fulton, R; van Ness, B (1994) Selective synergy of immunoglobulin enhancer elements in B-cell development: a characteristic of kappa light chain enhancers, but not heavy chain enhancers. Nucleic Acids Res 22:4216-23
Schanke, J T; Van Ness, B G (1994) Organization of the transcription factor binding sites in the kappa Ig intron enhancer. Effects of position, orientation, and spacing. J Immunol 153:4565-72
Billadeau, D; Ahmann, G; Greipp, P et al. (1993) The bone marrow of multiple myeloma patients contains B cell populations at different stages of differentiation that are clonally related to the malignant plasma cell. J Exp Med 178:1023-31
Zaknoen, S L; Christian, S L; Suen, R et al. (1992) B-chronic lymphocytic leukemia cells contain both endogenous kappa immunoglobulin mRNA and critical immunoglobulin gene activation transcription factors. Leukemia 6:675-9
Billadeau, D; Quam, L; Thomas, W et al. (1992) Detection and quantitation of malignant cells in the peripheral blood of multiple myeloma patients. Blood 80:1818-24
Martin, D; Huang, R Q; LeBien, T et al. (1991) Induced rearrangement of kappa genes in the BLIN-1 human pre-B cell line correlates with germline J-C kappa and V kappa transcription. J Exp Med 173:639-45
Billadeau, D; Blackstadt, M; Greipp, P et al. (1991) Analysis of B-lymphoid malignancies using allele-specific polymerase chain reaction: a technique for sequential quantitation of residual disease. Blood 78:3021-9

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