Elucidating IgH transcriptional regulation is critical for understanding how the humoral immune response is controlled. In addition, it provides an important paradigm for understanding tissue-specific and developmentally regulated gene transcription. The investigators past work has helped to identify and characterize proteins which regulate the transcriptional activity of VH promoters and the Eu enhancer. To continue studies on IgH transcriptional regulation, the investigator proposes experiments in three general areas: 1) to determine the biological importance of particular IgH transcription proteins in the context of B-cell development and function, 2) to identify the biochemical mechanisms of action of IgH transcription proteins and 3) to study the role of germline VH transcripts in VDJ recombination. Studies will focus primarily on C/EBP and uE3 binding proteins based on the investigator's past experience and because the play an important role in IgH transcription. To study the biological role of uE3 and C/EBP binding proteins, mice deficient in TFE3 or Ig/EBP will be created and studied. Long term bone marrow cultures will be exploited to study the roles of uE3 and C/EBP proteins in primary B-cell development and for lymphopoiesis in vivo. In vitro transcription utilizing reconstituted chromatin templates will be used to study the biochemical mechanism of action of uE3 and c/EBP proteins bound to both distal (enhancer) and proximal sites. RAG-/- cell lines will be studied to determine if germline VH transcripts are required for VDJ recombination and gene targeting will be employed to test whether transcription and/or altered chromatin structure are sufficient to target recombination of particular VH gene segments.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029361-20
Application #
2838471
Study Section
Special Emphasis Panel (ZRG2-IMS (03))
Program Officer
Tompkins, Laurie
Project Start
1988-12-01
Project End
2001-11-30
Budget Start
1998-12-01
Budget End
1999-11-30
Support Year
20
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
167204994
City
New York
State
NY
Country
United States
Zip Code
10032
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Lin, Kuo-I; Angelin-Duclos, Cristina; Kuo, Tracy C et al. (2002) Blimp-1-dependent repression of Pax-5 is required for differentiation of B cells to immunoglobulin M-secreting plasma cells. Mol Cell Biol 22:4771-80
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Angelin-Duclos, Cristina; Johnson, Kristen; Liao, Jerry et al. (2002) An interfering form of Blimp-1 increases IgM secreting plasma cells and blocks maturation of peripheral B cells. Eur J Immunol 32:3765-75
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Calame, K L (2001) Plasma cells: finding new light at the end of B cell development. Nat Immunol 2:1103-8
Piskurich, J F; Lin, K I; Lin, Y et al. (2000) BLIMP-I mediates extinction of major histocompatibility class II transactivator expression in plasma cells. Nat Immunol 1:526-32
Lin, K I; Lin, Y; Calame, K (2000) Repression of c-myc is necessary but not sufficient for terminal differentiation of B lymphocytes in vitro. Mol Cell Biol 20:8684-95
Berrier, A; Siu, G; Calame, K (1998) Transcription of a minimal promoter from the NF-IL6 gene is regulated by CREB/ATF and SP1 proteins in U937 promonocytic cells. J Immunol 161:2267-75
Angelin-Duclos, C; Calame, K (1998) Evidence that immunoglobulin VH-DJ recombination does not require germ line transcription of the recombining variable gene segment. Mol Cell Biol 18:6253-64

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