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Molecular and Cellular Biology, October 2003, p. 7055-7067, Vol. 23, No. 19
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.19.7055-7067.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of mRNAs Associated with {alpha}CP2-Containing RNP Complexes

Shelly A. Waggoner and Stephen A. Liebhaber*

Departments of Genetics and Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

Received 12 March 2003/ Returned for modification 10 June 2003/ Accepted 1 July 2003

Posttranscriptional controls in higher eukaryotes are central to cell differentiation and developmental programs. These controls reflect sequence-specific interactions of mRNAs with one or more RNA binding proteins. The {alpha}-globin poly(C) binding proteins ({alpha}CPs) comprise a highly abundant subset of K homology (KH) domain RNA binding proteins and have a characteristic preference for binding single-stranded C-rich motifs. {alpha}CPs have been implicated in translation control and stabilization of multiple cellular and viral mRNAs. To explore the full contribution of {alpha}CPs to cell function, we have identified a set of mRNAs that associate in vivo with the major {alpha}CP2 isoforms. One hundred sixty mRNA species were consistently identified in three independent analyses of {alpha}CP2-RNP complexes immunopurified from a human hematopoietic cell line (K562). These mRNAs could be grouped into subsets encoding cytoskeletal components, transcription factors, proto-oncogenes, and cell signaling factors. Two mRNAs were linked to ceroid lipofuscinosis, indicating a potential role for {alpha}CP2 in this infantile neurodegenerative disease. Surprisingly, {alpha}CP2 mRNA itself was represented in {alpha}CP2-RNP complexes, suggesting autoregulatory control of {alpha}CP2 expression. In vitro analyses of representative target mRNAs confirmed direct binding of {alpha}CP2 within their 3' untranslated regions. These data expand the list of mRNAs that associate with {alpha}CP2 in vivo and establish a foundation for modeling its role in coordinating pathways of posttranscriptional gene regulation.

* Corresponding author. Mailing address: Department of Genetics, University of Pennsylvania School of Medicine, Room 428, Clinical Research Building, 415 Curie Blvd., Philadelphia, PA 19104. Phone: (215) 898-7834. Fax: (215) 573-5157. E-mail: liebhabe{at}mail.med.upenn.edu.

Molecular and Cellular Biology, October 2003, p. 7055-7067, Vol. 23, No. 19
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.19.7055-7067.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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