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Eukaryotic Cell, May 2006, p. 806-815, Vol. 5, No. 5
1535-9778/06/$08.00+0     doi:10.1128/EC.5.5.806-815.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The SUMO Pathway Is Developmentally Regulated and Required for Programmed DNA Elimination in Paramecium tetraurelia{dagger} ,{ddagger}

Atsushi Matsuda and James D. Forney*

Department of Biochemistry, Purdue University, 175 S. University St., West Lafayette, Indiana 47907-2063

Received 22 October 2005/ Accepted 13 March 2006

Extensive genome-wide remodeling occurs during the formation of the somatic macronuclei from the germ line micronuclei in ciliated protozoa. This process is limited to sexual reproduction and includes DNA amplification, chromosome fragmentation, and the elimination of internal segments of DNA. Our efforts to define the pathways regulating these events revealed a gene encoding a homologue of ubiquitin activating enzyme 2 (UBA2) that is upregulated at the onset of macronuclear development in Paramecium tetraurelia. Uba2 enzymes are known to activate the protein called small ubiquitin-related modifier (SUMO) that is covalently attached to target proteins. Consistent with this relationship, Northern analysis showed increased abundance of SUMO transcripts during sexual reproduction in Paramecium. RNA interference (RNAi) against UBA2 or SUMO during vegetative growth had little effect on cell survival or fission rates. In contrast, RNAi of mating cells resulted in failure to form a functional macronucleus. Despite normal amplification of the genome, excision of internal eliminated sequences was completely blocked. Additional experiments showed that the homologous UBA2 and SUMO genes in Tetrahymena thermophila are also upregulated during conjugation. These results provide evidence for the developmental regulation of the SUMO pathway in ciliates and suggest a key role for the pathway in controlling genome remodeling.

* Corresponding author. Mailing address: Department of Biochemistry, Purdue University, 175 S. University St., West Lafayette, IN 47907-2063. Phone: (765) 494-1632. Fax: (765) 494-7897. E-mail: forney{at}purdue.edu.

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

{ddagger} This is paper number 2006-17876 from the Purdue Agriculture Experiment Station.

Eukaryotic Cell, May 2006, p. 806-815, Vol. 5, No. 5
1535-9778/06/$08.00+0     doi:10.1128/EC.5.5.806-815.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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