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Molecular and Cellular Biology, November 2002, p. 8079-8087, Vol. 22, No. 22
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.22.8079-8087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Human Pot1 (Protection of Telomeres) Protein: Cytolocalization, Gene Structure, and Alternative Splicing

Peter Baumann,{dagger} Elaine Podell, and Thomas R. Cech*

Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309-0215

Received 5 June 2002/ Returned for modification 18 July 2002/ Accepted 19 August 2002

Fission yeast Pot1 (protection of telomeres) is a single-stranded telomeric DNA binding protein with a critical role in ensuring chromosome stability. A putative human homolog (hPot1) was previously identified, based on moderate sequence similarity with fission yeast Pot1 and telomere end-binding proteins from ciliated protozoa. Using indirect immunofluorescence, we show here that epitope-tagged hPot1 localizes to telomeres in interphase nuclei of human cells, consistent with a direct role in telomere end protection. The hPOT1 gene contains 22 exons, most of which are present in all cDNAs examined. However, four exons are subject to exon skipping in some transcripts, giving rise to five splice variants. Four of these are ubiquitously expressed, whereas the fifth appears to be specific to leukocytes. The resultant proteins vary significantly in their ability to form complexes with single-stranded telomeric DNA as judged by electrophoretic mobility shift assays. In addition to these splice variants, the Pot1 family is expanded by the identification of six more genes from diverse species. Pot1-like proteins have now been found in plants, animals, yeasts, and microsporidia.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, Dept. of Chemistry & Biochemistry, University of Colorado, Boulder, CO 80309-0215. Phone: (303) 492-8606. Fax: (303) 492-6194. E-mail: thomas.cech{at}colorado.edu.

{dagger} Present address: Stowers Institute for Medical Research, Kansas City, MO 64110.

Molecular and Cellular Biology, November 2002, p. 8079-8087, Vol. 22, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.22.8079-8087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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