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doi: 10.1242/10.1242/jcs.00137

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RNA trafficking and stabilization elements associate with multiple brain proteins

Mark Snee^*, Grahame J. Kidd¹, Trent P. Munro² and Ross Smith

Department of Biochemistry and Molecular Biology, The University of Queensland, Qld 4072, Australia
¹ Department of Neurosciences NC30, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland OH 44195, USA
² Wellcome/CR UK Institute, Tennis Court Rd, Cambridge CB2 1QR, UK
^* Present address: Institute for Cellular and Molecular Biology, University of Texas, 2500 Speedway, Austin, TX, USA

Author for correspondence (e-mail: ross.s{at}uq.edu.au)

Accepted 3 September 2002

Two of the best understood somatic cell mRNA cytoplasmic trafficking elements are those governing localization of ß-actin and myelin basic protein mRNAs. These cis-acting elements bind the trans-acting factors fibroblast ZBP-1 and hnRNP A2, respectively. It is not known whether these elements fulfil other roles in mRNA metabolism. To address this question we have used Edman sequencing and western blotting to identify six rat brain proteins that bind the ß-actin element (zipcode). All are known RNA-binding proteins and differ from ZBP-1. Comparison with proteins that bind the hnRNP A2 and AU-rich response elements, A2RE/A2RE11 and AURE, showed that AURE and zipcode bind a similar set of proteins that does not overlap with those that bind A2RE11. The zipcode-binding protein, KSRP, and hnRNP A2 were selected for further study and were shown by confocal immunofluorescence microscopy to have similar distributions in the central nervous system, but they were found in largely separate locations in cell nuclei. In the cytoplasm of cultured oligodendrocytes they were segregated into separate populations of cytoplasmic granules. We conclude that not only may there be families of trans-acting factors for the same cis-acting element, which are presumably required at different stages of mRNA processing and metabolism, but independent factors may also target different and multiple RNAs in the same cell.

Key words: RNA-protein interactions, Zipcode, RNA trafficking, Protein sequencing, Confocal microscopy

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