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Molecular and Cellular Biology, November 2006, p. 8242-8251, Vol. 26, No. 22
0270-7306/06/$08.00+0     doi:10.1128/MCB.01348-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Role of RNA Polymerase III Transcription Factors in the Selection of Integration Sites by the Dictyostelium Non-Long Terminal Repeat Retrotransposon TRE5-A{triangledown}

Oliver Siol,1,2 Moustapha Boutliliss,1,2 Thanh Chung,1 Gernot Glöckner,3 Theodor Dingermann,1,4 and Thomas Winckler2*

Institut für Pharmazeutische Biologie, Universität Frankfurt/M., Frankfurt, Germany,1 Lehrstuhl für Pharmazeutische Biologie, Universität Jena, Jena, Germany,2 Genome Analysis, Fritz Lipmann Institute, Jena, Germany,3 Zentrum für Arzneimittelforschung, Entwicklung und Sicherheit, Frankfurt, Germany4

Received 24 July 2006/ Returned for modification 18 August 2006/ Accepted 3 September 2006

In the compact Dictyostelium discoideum genome, non-long terminal repeat (non-LTR) retrotransposons known as TREs avoid accidental integration-mediated gene disruption by targeting the vicinity of tRNA genes. In this study we provide the first evidence that proteins of a non-LTR retrotransposon interact with a target-specific transcription factor to direct its integration. We applied an in vivo selection system that allows for the isolation of natural TRE5-A integrations into a known genomic location upstream of tRNA genes. TRE5-A frequently modified the integration site in a way characteristic of other non-LTR retrotransposons by adding nontemplated extra nucleotides and generating small and extended target site deletions. Mutations within the B-box promoter of the targeted tRNA genes interfered with both the in vitro binding of RNA polymerase III transcription factor TFIIIC and the ability of TRE5-A to target these genes. An isolated B box was sufficient to enhance TRE5-A integration in the absence of a surrounding tRNA gene. The RNA polymerase III-transcribed ribosomal 5S gene recruits TFIIIC in a B-box-independent manner, yet it was readily targeted by TRE5-A in our assay. These results suggest a direct role of an RNA polymerase III transcription factor in the targeting process.

* Corresponding author. Mailing address: Universität Jena, Lehrstuhl für Pharmazeutische Biologie, Semmelweisstrasse 10, D-07743 Jena, Germany. Phone: 49-3641-949841. Fax: 49-3641-949842. E-mail: t.winckler{at}uni-jena.de.

{triangledown} Published ahead of print on 18 September 2006.

Molecular and Cellular Biology, November 2006, p. 8242-8251, Vol. 26, No. 22
0270-7306/06/$08.00+0     doi:10.1128/MCB.01348-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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