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Volumn 292, Issue 5519, 2001, Pages 1171-1175

Pot1, the putative telomere end-binding protein in fission yeast and humans

Author keywords

[No Author keywords available]

Indexed keywords

CHROMOSOMES; DNA; PROTOZOA; YEAST;

EID: 0035844082     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1060036     Document Type: Article
Times cited : (845)

References (42)
  • 3
    • 0343721193 scopus 로고    scopus 로고
    • note
    • The presence of 3′ single-stranded extensions in S. pombe was verified by hybridization of native genomic DNA with a telomeric probe before and after exonuclease 1 treatment (25).
  • 12
    • 0342416022 scopus 로고    scopus 로고
    • note
    • The O. nova α subunit by itself preferentially binds the 3′ terminus of a telomeric oligonucleotide, but also binds single-stranded telomeric DNA internally (11).
  • 16
    • 0033553536 scopus 로고    scopus 로고
    • J. D. Griffith et al., Cell 97, 503 (1999).
    • (1999) Cell , vol.97 , pp. 503
    • Griffith, J.D.1
  • 17
    • 0343721188 scopus 로고    scopus 로고
    • note
    • + with the kanamycin resistance gene as described (34).
  • 21
    • 0342850899 scopus 로고    scopus 로고
    • note
    • 4 (pH 8.0), 0.6 M NaCl, 10% glycerol, 0.2% Tween-20, and 5 mM β-mercaptoethanol] containing increasing concentrations of imidazole. Pot1p eluted around 90 mM imidazote. Pot1p-containing fractions were dialyzed against T buffer [50 mM tris-HCl (pH 8.0), 10% glycerol, 0.5 mM EDTA, and 0.5 mM dithiothreitol] containing 0.2 M KCl, and Pot1p was further purified on a Q-sepharose column (Pharmacia) using a linear gradient of KCl (0.2 to 1 M). The 22-kD Pot1p fragment was found in the flowthrough, whereas Pot1p eluted around 0.5 M KCl. The protein was dialyzed against T buffer plus 0.2 M KCl and stored at -80°C.
  • 22
    • 0343721182 scopus 로고    scopus 로고
    • note
    • Identical results were obtained when hemagglutinin-tagged Pot1p was expressed in S. pombe and cell-free extracts were used in bandshift assays (19).
  • 24
  • 25
    • 0343721174 scopus 로고    scopus 로고
    • See Science Online (www.sciencemag.org/cgi/ content/full/292/5519/1171/DC1).
    • Science
  • 26
  • 27
    • 0030745448 scopus 로고    scopus 로고
    • M. Meyerson et al., Cell 90, 785 (1997).
    • (1997) Cell , vol.90 , pp. 785
    • Meyerson, M.1
  • 29
    • 0343721166 scopus 로고    scopus 로고
    • note
    • Human POT1 was amplified by polymerase chain reaction from ovary cDNA and cloned into the pQE30 expression vector. The protein was purified over Ni-NTA resin under the same conditions as SpPot1p. The human protein eluted around 135 mM imidazole.
  • 32
    • 0342415998 scopus 로고    scopus 로고
    • note
    • Yet another state may involve a complex with heterogeneous nuclear ribonucleoproteins, several of which have properties consistent with a role in telomere maintenance (35-38).
  • 33
    • 0342850880 scopus 로고    scopus 로고
    • note
    • Pot1p might also bind the displaced single strand of a D-loop within a t-loop, although the protein does have a preference for binding terminal rather than internal repeats.
  • 42
    • 0343721140 scopus 로고    scopus 로고
    • We thank D. Lyons, G. Mellitzer, T. Nakamura, O. Peersen, V. Wood, and the members of the Cech laboratory for helpful discussions; D. King for mass spectroscopy; D. Baumann, K. Goodrich, Y. Han, and E. Podell for technical assistance; and T. Bryan, K. Friedman, and A. Zaug for critical reading of the manuscript. P.B. was supported in part by a Wellcome Prize Traveling Research Fellowship (grant 054549/Z/98/Z)
    • We thank D. Lyons, G. Mellitzer, T. Nakamura, O. Peersen, V. Wood, and the members of the Cech laboratory for helpful discussions; D. King for mass spectroscopy; D. Baumann, K. Goodrich, Y. Han, and E. Podell for technical assistance; and T. Bryan, K. Friedman, and A. Zaug for critical reading of the manuscript. P.B. was supported in part by a Wellcome Prize Traveling Research Fellowship (grant 054549/Z/98/Z).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.