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Molecular and Cellular Biology, June 2000, p. 4128-4134, Vol. 20, No. 11
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Novel Chromodomain Protein, Pdd3p, Associates with Internal Eliminated Sequences during Macronuclear Development in Tetrahymena thermophila

Mikhail A. Nikiforov,dagger Martin A. Gorovsky, and C. David Allis*

Department of Biology, University of Rochester, Rochester, New York 14627

Received 1 December 1999/Returned for modification 3 January 2000/Accepted 3 March 2000

Conversion of the germ line micronuclear genome into the genome of a somatic macronucleus in Tetrahymena thermophila requires several DNA rearrangement processes. These include (i) excision and subsequent elimination of several thousand internal eliminated sequences (IESs) scattered throughout the micronuclear genome and (ii) breakage of the micronuclear chromosomes into hundreds of DNA fragments, followed by de novo telomere addition to their ends. Chromosome breakage sequences (Cbs) that determine the sites of breakage and short regions of DNA adjacent to them are also eliminated. Both processes occur concomitantly in the developing macronucleus. Two stage-specific protein factors involved in germ line DNA elimination have been described previously. Pdd1p and Pdd2p (for programmed DNA degradation) physically associate with internal eliminated sequences in transient electron-dense structures in the developing macronucleus. Here, we report the purification, sequence analysis, and characterization of Pdd3p, a novel developmentally regulated, chromodomain-containing polypeptide. Pdd3p colocalizes with Pdd1p in the peripheral regions of DNA elimination structures, but is also found more internally. DNA cross-linked and immunoprecipitated with Pdd1p- or Pdd3p-specific antibodies is enriched in IESs, but not Cbs, suggesting that different protein factors are involved in elimination of these two groups of sequences.

* Corresponding author. Present address: Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22908. Phone: (804) 243-6048. Fax: (804) 924-5069. E-mail: allis{at}virginia.edu.

dagger Present address: Department of Molecular Biology, Lewis Thomas Lab, Princeton University, Princeton, NJ 08544.

Molecular and Cellular Biology, June 2000, p. 4128-4134, Vol. 20, No. 11
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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