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Molecular and Cellular Biology, March 2005, p. 2073-2084, Vol. 25, No. 6
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.6.2073-2084.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Bruton's Tyrosine Kinase Regulates Immunoglobulin Promoter Activation in Association with the Transcription Factor Bright

Jaya Rajaiya,1 Melissa Hatfield,1 Jamee C. Nixon,1,2 David J. Rawlings,3 and Carol F. Webb1,2*

Immunobiology and Cancer Research Program, Oklahoma Medical Research Foundation,1 Departments of Microbiology and Immunology and Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma,2 Departments of Pediatrics and Immunology, University of Washington School of Medicine, Seattle, Washington3

Received 6 May 2004/ Returned for modification 6 July 2004/ Accepted 13 December 2004

Bright (B-cell regulator of immunoglobulin heavy chain transcription) binding to immunoglobulin heavy chain loci after B-cell activation is associated with increased heavy chain transcription. Our earlier reports demonstrated that Bright coimmunoprecipitates with Bruton's tyrosine kinase (Btk) and that these proteins associate in a DNA-binding complex in primary B cells. B cells from immunodeficient mice with a mutation in Btk failed to produce stable Bright DNA-binding complexes. In order to determine if Btk is important for Bright function, a transcription activation assay was established and analyzed using real-time PCR technology. Cells lacking both Bright and Btk were transfected with Bright and/or Btk along with an immunoglobulin heavy chain reporter construct. Immunoglobulin gene transcription was enhanced when Bright and Btk were coexpressed. In contrast, neither Bright nor Btk alone led to activation of heavy chain transcription. Furthermore, Bright function required both Btk kinase activity and sequences within the pleckstrin homology domain of Btk. Bright was not appreciably phosphorylated by Btk; however, a third tyrosine-phosphorylated protein coprecipitated with Bright. Thus, the ability of Bright to enhance immunoglobulin transcription critically requires functional Btk.

* Corresponding author. Mailing address: Oklahoma Medical Research Foundation, Immunobiology and Cancer Research Program, 825 N.E. 13th St., Oklahoma City, OK 73104. Phone: (405) 271-7564. Fax: (405) 271-7128. E-mail: carol-webb{at}omrf.ouhsc.edu.

Molecular and Cellular Biology, March 2005, p. 2073-2084, Vol. 25, No. 6
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.6.2073-2084.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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