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J. Biol. Chem., Vol. 278, Issue 37, 34943-34951, September 12, 2003

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The Roles of Intersubunit Interactions in Exosome Stability^*

Antonio M. Estévez  , Ben Lehner ¶, Christopher M. Sanderson ¶, Thomas Ruppert  and Christine Clayton 

From the Zentrum für Molekulare Biologie, Heidelberg Universität, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany and the ^¶United Kingdom Medical Research Council Human Genome Mapping Project Resource Centre, Hinxton, Cambridge CB10 1SB, United Kingdom

In eukaryotes, at least 10 proteins associate in a 3'-5' exonuclease complex, the exosome, which is involved in the processing of many RNA species. A recent model for the exosome placed six RNase PH-related components in a hexameric ring core structure, with three S1 domain proteins associated with the ring surface. So far, however, this model lacks experimental support. Using a combination of RNA interference, complex affinity purification, and yeast two-hybrid approaches, we show here that the RNase PH homologues are important for maintenance of complex integrity. In contrast, the S1 domain proteins are not required for complex stability, although they are required for exosome function. Our results are partially consistent with the proposed model of the exosome, but indicate a different arrangement of the RNase PH proteins.

Received for publication, May 21, 2003

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AJ430833 and AJ516002, for TbEAP2 and TbEAP4 genes, respectively.

The protein sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with acession number(s) CAD23610 and CAD56698, respectively.

^* This work was supported by the Deutsche Forschungsgemeinschaft. 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.

To whom correspondence should be addressed. Present address: Instituto de Parasitología y Biomedicina "López-Neyra," CSIC, C/Ventanilla 11, 18001 Granada, Spain. Tel.: 34-958-805184; Fax: 34-958-203323; E-mail: aestevez{at}ipb.csic.es.

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