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Originally published In Press as doi:10.1074/jbc.M207352200 on September 24, 2002

J. Biol. Chem., Vol. 277, Issue 49, 46940-46949, December 6, 2002
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Molecular Basis for the Loss of CD28 Expression in Senescent T Cells*

Abbe N. VallejoDagger §, Ewa BrylDagger , Klaus Klarskov||**, Stephen Naylor||Dagger Dagger , Cornelia M. WeyandDagger , and Jörg J. GoronzyDagger

From the Dagger  Departments of Medicine and Immunology and the || Biomedical Mass Spectrometry and Functional Proteonomics Facility, Mayo Clinic, Rochester, Minnesota 55905

CD28null T cells are the most consistent biological indicator of the aging immune system in humans and are predictors of immunoincompetence in the elderly. The loss of CD28 is the result of an inoperative transcriptional initiator (INR), which consists of two nonoverlapping alpha  and beta  motifs that have distinct protein binding profiles but function as a unit. In CD28null T cells, there is a coordinate loss of alpha -/beta -bound complexes, hence the alpha beta -INR is inactive. In the present work therefore, studies were conducted to identify the components of such complexes that may account for the trans-activation of the alpha beta -INR. By affinity chromatography and tandem mass spectrometry, two proteins, namely, nucleolin and the A isoform of heterogeneous nuclear ribonucleoprotein-D0 (hnRNP-D0A), were identified to be among the key components of the site alpha  complex. In DNA binding assays, specific antibodies indicated their antigenic presence in alpha -bound complexes. Transcription assays showed that they are both required in the trans-activation of alpha beta -INR-driven DNA templates. Because CD28 is T cell-restricted, and nucleolin and hnRNP-D0A are ubiquitous proteins, these results support the notion that cell-specific functions can be regulated by commonly expressed proteins. The present data also provide evidence for INR-regulated transcription that is independent of the known components of the basal transcription complex.

* This work was supported by the Mayo Foundation, the Fogarty International Center, and National Institutes of Health Grants R01-AG15043, R01-AR41974, and R03-AR45830.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed: Guggenheim 401, Mayo Clinic, 200 First St. S.W., Rochester, MN 55905. Tel.: 507-284-1650; Fax: 507-284-5045; E-mail: vallejo.abbe@mayo.edu.

Present address: Dept. of Immunopathology, Medical University of Gdansk, Poland.

** Present address: Dépt. de Phamacologie, Université de Sherbrooke, Québec, Canada.

Dagger Dagger Present address: Beyond Genomics, Waltham, MA 02451.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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