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Eukaryotic Cell, August 2006, p. 1347-1359, Vol. 5, No. 8
1535-9778/06/$08.00+0     doi:10.1128/EC.00149-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Molecular Genetic Analysis of an SNF2/brahma-Related Gene in Tetrahymena thermophila Suggests Roles in Growth and Nuclear Development

Jeffrey S. Fillingham ,^, Jyoti Garg, Nora Tsao, Nama Vythilingum, Takamitsu Nishikawa, and Ronald E. Pearlman^*

Department of Biology, York University, Toronto, Ontario, Canada M3J 1P3

Received 24 May 2006/ Accepted 9 June 2006

We used a reverse genetic approach to identify three members of the SNF2 superfamily of chromatin remodeling genes in the ciliated protozoan Tetrahymena thermophila in order to investigate possible functions of ATP-dependent chromatin remodeling factors in growth and nuclear development. Comparative sequence analysis of the gene product of the Tetrahymena brahma-related gene (TtBRG1) indicates it is a member of the SNF2/BRM subgroup of the SNF2 superfamily. Northern analysis suggests that TtBRG1 has roles in growth and nuclear development in Tetrahymena. Indirect immunofluorescence analysis during nuclear development indicates that TtBrg1p localizes to both the parental and developing macronucleus of Tetrahymena during the time period corresponding to genome rearrangements. We generated germ line knockout heterokaryons for TtBRG1 and demonstrated that expression of the gene is required to complete nuclear development of Tetrahymena. In addition, the formation of distinct Pdd1p-containing structures is disturbed during the late stages of conjugation in TtBRG1 germ line knockout heterokaryons. We discuss these results in light of possible roles of SNF2-related proteins in growth and nuclear development of Tetrahymena.

Supplemental material for this article may be found at http://ec.asm.org/.

Present address: Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada.

These authors contributed equally to this work.