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Molecular and Cellular Biology, November 2008, p. 6620-6631, Vol. 28, No. 21
0270-7306/08/$08.00+0     doi:10.1128/MCB.00448-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

TRF4 Is Involved in Polyadenylation of snRNAs in Drosophila melanogaster{triangledown}

Ryoichi Nakamura,1 Ryo Takeuchi,1 Kei-ichi Takata,1,2 Kaori Shimanouchi,1 Yoko Abe,1 Yoshihiro Kanai,1 Tatsushi Ruike,1 Ayumi Ihara,1 and Kengo Sakaguchi1*

Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi Chiba-ken, 278-8510, Japan,1 Department of Pharmacology, Hillman Cancer Center, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania 15213-18632

Received 19 March 2008/ Returned for modification 24 April 2008/ Accepted 20 August 2008

The Saccharomyces cerevisiae poly(A) polymerases Trf4 and Trf5 are involved in an RNA quality control mechanism, where polyadenylated RNAs are degraded by the nuclear exosome. Although Trf4/5 homologue genes are distributed throughout multicellular organisms, their biological roles remain to be elucidated. We isolated here the two homologues of Trf4/5 in Drosophila melanogaster, named DmTRF4-1 and DmTRF4-2, and investigated their biological function. DmTRF4-1 displayed poly(A) polymerase activity in vitro, whereas DmTRF4-2 did not. Gene knockdown of DmTRF4-1 by RNA interference is lethal in flies, as is the case for the trf4 trf5 double mutants. In contrast, disruption of DmTRF4-2 results in viable flies. Cellular localization analysis suggested that DmTRF4-1 localizes in the nucleolus. Abnormal polyadenylation of snRNAs was observed in transgenic flies overexpressing DmTRF4-1 and was slightly increased by the suppression of DmRrp6, the 3'-5' exonuclease of the nuclear exosome. These results suggest that DmTRF4-1 and DmRrp6 are involved in the polyadenylation-mediated degradation of snRNAs in vivo.

* Corresponding author. Mailing address: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi Chiba-ken, 278-8510, Japan. Phone: 81 4 7124 1501, ext. 3409. Fax: 81 4 7123 9767. E-mail: kengo{at}rs.noda.tus.ac.jp

{triangledown} Published ahead of print on 2 September 2008.

Molecular and Cellular Biology, November 2008, p. 6620-6631, Vol. 28, No. 21
0270-7306/08/$08.00+0     doi:10.1128/MCB.00448-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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