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J. Biol. Chem., Vol. 279, Issue 10, 8723-8731, March 5, 2004

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Annexin A2 Is a Novel RNA-binding Protein

Nolan R. Filipenko, Travis J. MacLeod, Chang-Soon Yoon, and David M. Waisman

From the Cancer Biology Research Group, Departments of Biochemistry & Molecular Biology and Oncology, University of Calgary, Calgary, Alberta T2N 4N1, Canada

Annexin A2 (ANXA2) is a Ca²⁺-binding protein that is up-regulated in virally transformed cell lines and in human tumors. Here, we show that ANXA2 binds directly to both ribonucleotide homopolymers and human c-myc RNA. ANXA2 was shown to bind specifically to poly(G) with high affinity (K_d = 60 nM) and not to poly(A), poly(C), or poly(U). The binding of ANXA2 to poly(G) required Ca²⁺ (A_50% = 10 µM). The presence of RNA in the immunoprecipitates of ANXA2 isolated from HeLa cells established that ANXA2 formed a ribonucleoprotein complex in vivo. Sucrose gradient analysis showed that ANXA2 associates with ribonucleoprotein complexes and not with polyribosomes. Reverse transcriptase-PCR identified c-myc mRNA as a component of the ribonucleoprotein complex formed by ANXA2 in vivo, and binding studies confirmed a direct interaction between ANXA2 and c-myc mRNA. Transfection of LNCaP cells with the ANXA2 gene resulted in the up-regulation of c-Myc protein. These findings identify ANXA2 as a Ca²⁺-dependent RNA-binding protein that interacts with the mRNA of the nuclear oncogene, c-myc.

Received for publication, October 31, 2003 , and in revised form, December 4, 2003.

* This work was supported by National Institutes of Health Grant RO1CA 78639 and grants from the Alberta Heart and Stroke Foundation and the Canadian Institutes of Health Research. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by studentships from the Alberta Heritage Foundation for Medical Research and the Alberta Cancer Board.

To whom correspondence should be addressed: Depts. of Biochemistry & Molecular Biology and Oncology, Faculty of Medicine, University of Calgary, 3330 Hospital Dr. N.W., Calgary, Alberta T2N 4N1, Canada. Tel.: 403-220-3022; Fax: 403-283-4841; E-mail: waisman{at}ucalgary.ca.

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