The rearrangement of immunoglobulin (and T cell receptor) genes is at many levels. Developmentally, in the B cell lineage, only proB and preB cells have the VDJ recombinase. The rearrangement substrate must be accessible, which presumably means it must be in an open chromatin configuration. Rearrangement signal sequences flanking the V,D and J gene joining partners must have the correct consensus sequence. Thus, one has some general picture of the rearrangement process, but many of the details of the molecular mechanism and its regulation are unknown.
The specific aims of this proposal are to analyze in depth the requirements of immunoglobulin gene targets for rearrangement and the conditions for a correct rearrangement process in preB cells. The following experiments are planned: 1) Transgenic mice which carry a rearrangement test transgene will be studied to determine how the chromatin status of the transgene relates to its accessibility for correct rearrangement. This will include mapping a control gene(s) (the """"""""strain specific modifier"""""""", SSM-1) which determines methylation and non-rearrangement in several transgenic lines and cloning of the SSM-1 gene; analysis of hybridomas and preB cell lines from the transgenic mice to explain the relationship between methylation of the transgene and its immunoglobulin gene specific rearrangement. 2) Determination of the effect of target gene methylation in vitro on immunoglobulin gene rearrangement. 3) Investigation of a specific suppressor of immunoglobulin gene rearrangement. 4) Study of the expression and function of a gene (p61) which encodes a DNA binding protein, possibly involved in immunoglobulin gene rearrangement. All the planned experiments will aid in the understanding of the expression and function of the partner genes involved in immunoglobulin gene rearrangement. Understanding this mechanism will provide a fundamental basis for the unraveling of many diseases involving the immune system, such as autoimmunities, infectious diseases, allergies and cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AI024780-11
Application #
2003421
Study Section
Special Emphasis Panel (NSS)
Project Start
1986-09-30
Project End
2001-11-30
Budget Start
1996-12-01
Budget End
1997-11-30
Support Year
11
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Chicago
Department
Genetics
Type
Schools of Medicine
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
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Sun, T; Ezekiel, U R; Erskine, L et al. (1999) Signal joint formation is inhibited in murine scid preB cells and fibroblasts in substrates with homopolymeric coding ends. Mol Immunol 36:551-8
Engler, P; Storb, U (1999) Hypomethylation is necessary but not sufficient for V(D)J recombination within a transgenic substrate. Mol Immunol 36:1169-73
Engler, P; Doglio, L T; Bozek, G et al. (1998) A cis-acting element that directs the activity of the murine methylation modifier locus Ssm1. Proc Natl Acad Sci U S A 95:10763-8
Ezekiel, U R; Sun, T; Bozek, G et al. (1997) The composition of coding joints formed in V(D)J recombination is strongly affected by the nucleotide sequence of the coding ends and their relationship to the recombination signal sequences. Mol Cell Biol 17:4191-7
Fuller, K; Storb, U (1997) Identification and characterization of the murine Rag1 promoter. Mol Immunol 34:939-54
Weng, A; Magnuson, T; Storb, U (1995) Strain-specific transgene methylation occurs early in mouse development and can be recapitulated in embryonic stem cells. Development 121:2853-9
Weng, A; Engler, P; Storb, U (1995) The bulk chromatin structure of a murine transgene does not vary with its transcriptional or DNA methylation status. Mol Cell Biol 15:572-9
Storb, U; Roth, P; Kurtz, B K (1994) Gamma 2b transgenic mice as a model for the role of immunoglobulins in B cell development. Immunol Res 13:291-8
Engler, P; Weng, A; Storb, U (1993) Influence of CpG methylation and target spacing on V(D)J recombination in a transgenic substrate. Mol Cell Biol 13:571-7

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