Protection of internal (TTAGGG)n repeats in Chinese hamster cells by telomeric protein TRF1.
Chinese hamster cells have large interstitial (TTAGGG) bands (ITs) which are unstable and should be protected by an unknown mechanism. Here, we expressed in Chinese hamster V79 cells green fluorescent protein (GFP)-tagged human TRF1, and found that a major fraction of GFP-TRF1 bound to ITs is diffusionally mobile. This fraction strongly decreases after treatment of cells with wortmannin, a protein kinase inhibitor, and this drug also increases the frequency of chromosome aberrations. Ionizing radiation does not induce detectable translocation of GFP-TRF1 to the sites of random double-strand breaks visualized using antibodies against histone gamma-H2AX. TRF1 is known to be eliminated from telomeres by overexpression of tankyrase 1 which induces TRF1 poly(ADP-ribosyl)ation. We transfected V79 cells by plasmid encoding tankyrase 1 and found that the frequency of chromosome rearrangements is increased in these cells independently of their treatment by IR. Taken together, our results suggest that TRF1 is involved in sequence-specific protection of internal nontelomeric (TTAGGG)n repeats.
Krutilina
Raisa Ivanovna
RI
Laboratory of Chromosome Stability, Institute of Cytology, Russian Academy of Sciences, 194064 St Petersburg, Russia.
Smirnova
Alexandra Nikolaevna
AN
Mudrak
Olga Stanislavovna
OS
Pleskach
Nadezhda Mikhailovna
NM
Svetlova
Maria Pavlovna
MP
Oei
Shiao-Li
SL
Yau
Peter M
PM
Bradbury
Edwin Morton
EM
Zalensky
Andrey Olegovich
AO
Tomilin
Nikolai Viktorovich
NV
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001761:Bleomycin / Q000494:pharmacology; D002460:Cell Line; D002869:Chromosome Aberrations; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006657:Histones / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D038501:Tankyrases / Q000096:biosynthesis; D016615:Telomere / Q000378:metabolism*; D035321:Telomeric Repeat Binding Protein 1 / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D014162:Transfection; D000077191:Wortmannin
D000730:Androstadienes; D001761:Bleomycin; D004791:Enzyme Inhibitors; D049452:Green Fluorescent Proteins; D006657:Histones; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D038501:Tankyrases; D035321:Telomeric Repeat Binding Protein 1; D000077191:Wortmannin; C418109:TNKS protein, human