title: Loss of hPot1 function leads to telomere instability and a cut-like phenotype.
abstract: The human telomere binding protein hPot1 binds to the most distal single-stranded extension of telomeric DNA in vitro, and probably in vivo, as well as associating with the double-stranded telomeric DNA binding proteins TRF1 and TRF2 through the bridging proteins PTOP (also known as PIP1 or TINT1) and TIN2. Disrupting either the DNA binding activity of hPot1 or its association with PTOP results in elongated telomeres, suggesting a role for hPot1 in telomere length regulation. However, mutations to POT1 and Cdc13p, the fission and budding yeast genes encoding the structural orthologs of this protein, leads to telomere instability and cell death. Thus, it is possible that the hPot1 protein may also serve to cap and protect telomeres in humans. Indeed, we now find that knocking down the expression of hPot1 in human cells causes apoptosis or senescence, as well as an increase in telomere associations and anaphase bridges, telltale signs of telomere instability. In addition, knockdown cells also displayed chromatin bridges between interphase cells, reminiscent of the cut phenotype that was first described in fission yeast and in which cytokinesis progresses despite a failure of chromatid separation. However, unlike the yeast cut phenotypes, we suggest that the cut-like phenotype observed in hPot1 knockdown cells is a consequence of the fusion of chromosome ends and that this fusion impedes proper chromosomal segregation. We conclude that hPot1 protects chromosome ends from illegitimate recombination, catastrophic chromosome instability, and abnormal chromosome segregation.
authors.affiliation: Department of Pharmacology and Cancer Biology, Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Veldman
authors.forename: Katherine T
authors.initials: KT
authors.lastname: Etheridge
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Counter
mesh_terms: D017209:Apoptosis / Q000502:physiology; D043171:Chromosomal Instability / Q000235:genetics / Q000502:physiology*; D020090:Chromosome Segregation / Q000502:physiology; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D048629:Micronuclei, Chromosome-Defective; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000089804:Shelterin Complex; D016615:Telomere / Q000235:genetics / Q000502:physiology*; D034501:Telomere-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D035321:Telomeric Repeat Binding Protein 1 / Q000378:metabolism; D035341:Telomeric Repeat Binding Protein 2 / Q000378:metabolism
chemical_list: C495876:ACD protein, human; D017931:DNA Primers; C440849:POT1 protein, human; D000089804:Shelterin Complex; C440868:TINF2 protein, human; D034501:Telomere-Binding Proteins; D035321:Telomeric Repeat Binding Protein 1; D035341:Telomeric Repeat Binding Protein 2; D004247:DNA