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Molecular and Cellular Biology, June 2006, p. 4499-4510, Vol. 26, No. 12
0270-7306/06/$08.00+0     doi:10.1128/MCB.00079-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Centromeric Histone H3 Is Essential for Vegetative Cell Division and for DNA Elimination during Conjugation in Tetrahymena thermophila{dagger}

Bowen Cui and Martin A. Gorovsky*

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

Received 12 January 2006/ Returned for modification 16 March 2006/ Accepted 3 April 2006

The Tetrahymena thermophila CNA1 gene encodes the centromeric H3, Cna1p. Green fluorescent protein (GFP)-tagged Cna1p localizes in micronuclei in dots whose number and behavior during mitosis and conjugation are consistent with centromeres. During interphase, Cna1p-GFP localizes in peripheral dots, suggesting centromeres are associated with the nuclear envelope. Newly synthesized Cna1p-GFP enters micronuclei in mitosis and accumulates in the nucleoplasm. Its deposition at centromeres starts at early S phase and continues through most of S phase. CNA1 is required for vegetative cell growth. Knockdown of CNA1 genes in the somatic macronucleus results in micronuclear DNA loss and delayed chromosome segregation during mitosis. During conjugation, Cna1p-GFP disappears from the centromeres in the developing macronucleus, consistent with centromeric sequences being internal eliminated sequences. Surprisingly, zygotic CNA1 is required for efficient elimination of germ line-specific sequences during development of the new macronuclei but not for the RNA interference pathway, through which sequences are targeted for elimination. Zygotically expressed Cna1p localizes in the spherical structures in which the later stages of DNA elimination occur, and these structures cannot be formed in the absence of zygotic CNA1, suggesting that, in addition to functioning in centromeres, Cna1p may also play a role in organizing the formation of the DNA elimination structures.

* Corresponding author. Mailing address: Department of Biology, University of Rochester, Rochester, NY 14627. Phone: (585) 275-6988. Fax: (585) 275-2070. E-mail: goro{at}mail.rochester.edu.

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

Molecular and Cellular Biology, June 2006, p. 4499-4510, Vol. 26, No. 12
0270-7306/06/$08.00+0     doi:10.1128/MCB.00079-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Loidl, J., Mochizuki, K. (2009). Tetrahymena Meiotic Nuclear Reorganization Is Induced by a Checkpoint Kinase-dependent Response to DNA Damage. Mol. Biol. Cell 20: 2428-2437 [Abstract] [Full Text]  
  • Aronica, L., Bednenko, J., Noto, T., DeSouza, L. V., Siu, K.W. M., Loidl, J., Pearlman, R. E., Gorovsky, M. A., Mochizuki, K. (2008). Study of an RNA helicase implicates small RNA-noncoding RNA interactions in programmed DNA elimination in Tetrahymena. Genes Dev. 22: 2228-2241 [Abstract] [Full Text]  
  • Mochizuki, K., Novatchkova, M., Loidl, J. (2008). DNA double-strand breaks, but not crossovers, are required for the reorganization of meiotic nuclei in Tetrahymena. J. Cell Sci. 121: 2148-2158 [Abstract] [Full Text]  
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