This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Anzai, T.
Right arrow Articles by Fujiwara, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Anzai, T.
Right arrow Articles by Fujiwara, H.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, January 2001, p. 100-108, Vol. 21, No. 1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.1.100-108.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Sequence-Specific Recognition and Cleavage of Telomeric Repeat (TTAGG)n by Endonuclease of Non-Long Terminal Repeat Retrotransposon TRAS1

Tomohiro Anzai, Hidekazu Takahashi, and Haruhiko Fujiwara*

Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan

Received 13 March 2000/Returned for modification 4 May 2000/Accepted 13 October 2000

The telomere of the silkworm Bombyx mori consists of (TTAGG/CCTAA)n repeats and harbors a large number of telomeric repeat-specific non-long terminal repeat retrotransposons, such as TRAS1 and SART1. To understand how these retrotransposons recognize and integrate into the telomeric repeat in a sequence-specific manner, we expressed the apurinic-apryrimidinic endonuclease-like endonuclease domain of TRAS1 (TRAS1 EN), which is supposed to digest the target DNA, and characterized its enzymatic properties. Purified TRAS1 EN could generate specific nicks on both strands of the telomeric repeat sequence between T and A of the (TTAGG)n strand (bottom strand) and between C and T of the (CCTAA)n strand (top strand). These sites are consistent with insertion sites expected from the genomic structure of boundary regions of TRAS1. Time course studies of nicking activities on both strands revealed that the cleavages on the bottom strand preceded those on the top strand, supporting the target-primed reverse transcription model. TRAS1 EN could cleave the telomeric repeats specifically even if it was flanked by longer tracts of nontelomeric sequence, indicating that the target site specificity of the TRAS1 element was mainly determined by its EN domain. Based on mutation analyses, TRAS1 EN recognizes less than 10 bp around the initial cleavage site (upstream 7 bp and downstream 3 bp), and the GTTAG sequence especially is essential for the cleavage reaction on the bottom strand (5'. . . TTAGGTT down-arrow  AGG . . . 3'). TRAS1 EN, the first identified endonuclease digesting telomeric repeats, may be used as a genetic tool to shorten the telomere in insects and some other organisms.

* Corresponding author. Mailing address: Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan. Phone and Fax: 81-3-5841-4447. E-mail: haruh{at}k.u-tokyo.ac.jp.

Molecular and Cellular Biology, January 2001, p. 100-108, Vol. 21, No. 1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.1.100-108.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Maita, N., Aoyagi, H., Osanai, M., Shirakawa, M., Fujiwara, H. (2007). Characterization of the sequence specificity of the R1Bm endonuclease domain by structural and biochemical studies. Nucleic Acids Res 35: 3918-3927 [Abstract] [Full Text]  
  • Christensen, S. M., Bibillo, A., Eickbush, T. H. (2005). Role of the Bombyx mori R2 element N-terminal domain in the target-primed reverse transcription (TPRT) reaction. Nucleic Acids Res 33: 6461-6468 [Abstract] [Full Text]  
  • Kojima, K. K., Fujiwara, H. (2005). Long-Term Inheritance of the 28S rDNA-Specific Retrotransposon R2. Mol Biol Evol 22: 2157-2165 [Abstract] [Full Text]  
  • Christensen, S. M., Eickbush, T. H. (2005). R2 Target-Primed Reverse Transcription: Ordered Cleavage and Polymerization Steps by Protein Subunits Asymmetrically Bound to the Target DNA. Mol. Cell. Biol. 25: 6617-6628 [Abstract] [Full Text]  
  • Kojima, K. K., Fujiwara, H. (2005). An extraordinary retrotransposon family encoding dual endonucleases. Genome Res 15: 1106-1117 [Abstract] [Full Text]  
  • Hood, M. E., Katawczik, M., Giraud, T. (2005). Repeat-Induced Point Mutation and the Population Structure of Transposable Elements in Microbotryum violaceum. Genetics 170: 1081-1089 [Abstract] [Full Text]  
  • Anzai, T., Osanai, M., Hamada, M., Fujiwara, H. (2005). Functional roles of 3'-terminal structures of template RNA during in vivo retrotransposition of non-LTR retrotransposon, R1Bm. Nucleic Acids Res 33: 1993-2002 [Abstract] [Full Text]  
  • Maita, N., Anzai, T., Aoyagi, H., Mizuno, H., Fujiwara, H. (2004). Crystal Structure of the Endonuclease Domain Encoded by the Telomere-specific Long Interspersed Nuclear Element, TRAS1. J. Biol. Chem. 279: 41067-41076 [Abstract] [Full Text]  
  • Fujimoto, H., Hirukawa, Y., Tani, H., Matsuura, Y., Hashido, K., Tsuchida, K., Takada, N., Kobayashi, M., Maekawa, H. (2004). Integration of the 5' end of the retrotransposon, R2Bm, can be complemented by homologous recombination. Nucleic Acids Res 32: 1555-1565 [Abstract] [Full Text]  
  • Bringaud, F., Biteau, N., Zuiderwijk, E., Berriman, M., El-Sayed, N. M., Ghedin, E., Melville, S. E., Hall, N., Baltz, T. (2004). The ingi and RIME non-LTR Retrotransposons Are Not Randomly Distributed in the Genome of Trypanosoma brucei. Mol Biol Evol 21: 520-528 [Abstract] [Full Text]  
  • Matsumoto, T., Takahashi, H., Fujiwara, H. (2004). Targeted Nuclear Import of Open Reading Frame 1 Protein Is Required for In Vivo Retrotransposition of a Telomere-Specific Non-Long Terminal Repeat Retrotransposon, SART1. Mol. Cell. Biol. 24: 105-122 [Abstract] [Full Text]  
  • Kojima, K. K., Fujiwara, H. (2003). Evolution of Target Specificity in R1 Clade Non-LTR Retrotransposons. Mol Biol Evol 20: 351-361 [Abstract] [Full Text]  
  • Arkhipova, I. R., Morrison, H. G. (2001). From the Cover: Three retrotransposon families in the genome of Giardia lamblia: Two telomeric, one dead. Proc. Natl. Acad. Sci. USA 98: 14497-14502 [Abstract] [Full Text]  
  • Kubo, Y., Okazaki, S., Anzai, T., Fujiwara, H. (2001). Structural and Phylogenetic Analysis of TRAS, Telomeric Repeat-Specific Non-LTR Retrotransposon Families in Lepidopteran Insects. Mol Biol Evol 18: 848-857 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»