The Journal of Immunology, Vol 150, Issue 10 4376-4385, Copyright © 1993 by American Association of Immunologists

ARTICLES

Functional importance of the cyclic AMP response element-like decamer motif in the CD8 alpha promoter

MH Gao and PB Kavathas
Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06510.

Expression of the CD8 gene is highly regulated during lymphocyte differentiation and in a tissue-specific manner. We characterized the human CD8 alpha promoter region to determine whether tissue specificity resides within the promoter and to define important regulatory elements. A set of six fragments 5' of the CD8 alpha gene were linked to a luciferase reporter gene. The luciferase activity was then measured in a transient transfection assay. We found that CD8 alpha promoter activity can be detected from a 146-bp fragment upstream of the translation start site, but not from a 133-bp fragment. The cyclic AMP response element (CRE)-like site within the 10 bp from -143 to -133 is critical for promoter activity. Mutation of the CRE/decamer in the context of a 429-bp fragment causes loss of activity. Tissue specificity does not reside in the 146-bp fragment because this fragment directs expression in both T and non-T cell lines. Fragments longer than 146 bp are generally expressed less well in the cell lines suggesting the potential existence of negative regulatory elements upstream of -146. Using a CRE/decamer-containing oligomer as a probe in an electrophoresis mobility shift assay, three retarded bands formed by proteins binding to the DNA were detected using nuclear extracts from two T cell lines. Two of the three bands contain proteins of the CRE- binding protein (CREB)/activating transcription factor (ATF) family. Because the CRE-binding protein/activating transcription factor proteins play a role in the expression of many other T cell-specific genes, our work strengthens the hypothesis that the CRE motif is important for regulating the expression of T cell-specific genes.

This article has been cited by other articles:

				G. C. Grady, S. M. Mason, J. Stephen, J. C. Zuniga-Pflucker, and A. M. Michie Cyclic Adenosine 5'-Monophosphate Response Element Binding Protein Plays a Central Role in Mediating Proliferation and Differentiation Downstream of the Pre-TCR Complex in Developing Thymocytes J. Immunol., August 1, 2004; 173(3): 1802 - 1810. [Abstract] [Full Text] [PDF]

				L. Devine, L. J. Kieffer, V. Aitken, and P. B. Kavathas Human CD8{beta}, But Not Mouse CD8{beta}, Can Be Expressed in the Absence of CD8{alpha} as a {beta}{beta} Homodimer J. Immunol., January 15, 2000; 164(2): 833 - 838. [Abstract] [Full Text] [PDF]

				M. M. Khodadoust, K. D. Khan, and A. L. M. Bothwell Complex Regulation of Ly-6E Gene Transcription in T Cells by IFNs J. Immunol., July 15, 1999; 163(2): 811 - 819. [Abstract] [Full Text] [PDF]

				L. Devine, J. Sun, M. R. Barr, and P. B. Kavathas Orientation of the Ig Domains of CD8{alpha}{beta} Relative to MHC Class I J. Immunol., January 15, 1999; 162(2): 846 - 851. [Abstract] [Full Text] [PDF]

				M. M. Khodadoust, K. D. Khan, E.-h. Park, and A. L.M. Bothwell Distinct Regulatory Mechanisms for Interferon-alpha /beta (IFN-alpha /beta )- and IFN-gamma -Mediated Induction of Ly-6E Gene in B Cells Blood, October 1, 1998; 92(7): 2399 - 2409. [Abstract] [Full Text] [PDF]

				X. Deng, G.-R. Sun, Q. Zheng, and Y. Li Characterization of Human TCR Vbeta Gene Promoter. ROLE OF THE DODECAMER MOTIF IN PROMOTER ACTIVITY J. Biol. Chem., September 11, 1998; 273(37): 23709 - 23715. [Abstract] [Full Text] [PDF]

				X.-L. Zhang, R. Seong, R. Piracha, M. Larijani, M. Heeney, J. R. Parnes, and J. W. Chamberlain Distinct Stage-Specific cis-Active Transcriptional Mechanisms Control Expression of T Cell Coreceptor CD8{alpha} at Double- and Single-Positive Stages of Thymic Development J. Immunol., September 1, 1998; 161(5): 2254 - 2266. [Abstract] [Full Text] [PDF]

				R. Firestein and N. Feuerstein Association of Activating Transcription Factor 2 (ATF2) with the Ubiquitin-conjugating Enzyme hUBC9. IMPLICATION OF THE UBIQUITIN/PROTEASOME PATHWAY IN REGULATION OF ATF2 IN T CELLS J. Biol. Chem., March 6, 1998; 273(10): 5892 - 5902. [Abstract] [Full Text] [PDF]