The gene segments encoding immunoglobulins have the novel property of only being functionally expressed after they undergo specific genetic rearrangements in the developing B lymphocyte. In order to understand the complex controls which regulate antibody production it is necessary to examine the mechanisms by which the gene segments are rearranged and the elements which regulate their transcription. The formation of antibody genes requires site specific translocations of DNA, which for mouse Kappa light chains brings one of several hundred variable segments to one of four joining segments located about 3 kb upstream of a constant segment. Both functional and nonfunctional fragments of the rearranged Kappa locus in plasmacytomas, as well as reciprocal fragments which are generated by the rearrangement, will be characterized by DNA sequencing and compared to determine the consequence of Kappa rearrangements and structural requirements for transcription. The chromosomal location of Kappa rearrangements and origins of non Kappa DNA segments which rearrange into the Kappa locus will be determined. A systematic analysis of rearrangement will be followed in a clonally derived lymphoid cell line which rearranges the Kappa locus in culture. The elements which contribute to transcriptional efficiency will be determined by analyzing the transcriptional competence of aberrant rearrangements which represent mutations or deletions of the Kappa transcription unit. Studies will be initiated to identify specific regions of the DNA required for transcription in vivo by transient expression of Kappa genes in an SV40 vector-HeLa cell system. The ultimate goal of transcription studies will be to determine structures which affect interactions of DNA with elements important for transcriptional regulation in cell lines which can be stimulated or suppressed for antibody production.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
9R01GM037687-04
Application #
3293227
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1986-04-01
Project End
1991-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Iowa
Department
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
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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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