메뉴 건너뛰기




Volumn 276, Issue 5313, 1997, Pages 806-809

Cell cycle-dependent establishment of a late replication program

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; CELL CYCLE; DNA REPLICATION; NONHUMAN; PRIORITY JOURNAL; SACCHAROMYCES CEREVISIAE; TELOMERE;

EID: 0031005357     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.276.5313.806     Document Type: Article
Times cited : (141)

References (36)
  • 4
    • 0023440934 scopus 로고
    • R. S. Hansen, T. K. Canfield, M. M. Lamb, S. M. Gartler, C. D. Laird, Cell 73, 1403 (1993); C. D. Laird, Genetics 117, 587 (1987).
    • (1987) Genetics , vol.117 , pp. 587
    • Laird, C.D.1
  • 9
    • 0029300102 scopus 로고
    • J. F. Diffley, J. H. Cocker, S. J. Dowell, J. Harwood, A. Rowley, J. Cell Sci. Suppl. 19, 67 (1995); J. J. Li, Curr. Biol. 5, 472 (1995).
    • (1995) Curr. Biol. , vol.5 , pp. 472
    • Li, J.J.1
  • 15
    • 0026532151 scopus 로고
    • GAL-recombinase [made by deleting the 2-μm autonomously replicating sequence from plasmid pHM153 (14)]. One copy of the 65-bp recombinase target site [H. Araki et al., J. Mol. Biol. 225, 25 (1992)] was integrated in the "R14" fragment (5), ∼50 kb from the right end of chromosome V, with TRP1 as the selectable marker; the other copy of the recombinase site was integrated at the EcoR I site at the "R4/R5" junction, ∼20 kb from the telomere with URA3 as the marker. Upon excision of the cassette, the TRP1 marker is left behind on the chromosome while URA3 is retained on the circle. For the experiment on long-term growth of cells with the excised circle, cells were plated on minimal medium lacking uracil after induction of recombination. Clones with the excised circle were identified by CHEF gel electrophoresis and by Southern blotting, probing for the expected junction fragment on the circle. To overcome poor growth arising from loss of the circle (which contains essential genes like RAD3), we integrated a centromere (CEN5) in the URA3 gene on the circle. Integrants were selected by growth on plates containing 5-fluoroorotic acid, which selects against expression of URA3 [J. D. Boeke, F. LaCroute, G. R. Fink, Mol. Gen. Genet. 197, 345 (1984)], and correct integration of the centromere fragment was confirmed by Southern blotting and hybridization.
    • (1992) J. Mol. Biol. , vol.225 , pp. 25
    • Araki, H.1
  • 16
    • 0021668558 scopus 로고
    • GAL-recombinase [made by deleting the 2-μm autonomously replicating sequence from plasmid pHM153 (14)]. One copy of the 65-bp recombinase target site [H. Araki et al., J. Mol. Biol. 225, 25 (1992)] was integrated in the "R14" fragment (5), ∼50 kb from the right end of chromosome V, with TRP1 as the selectable marker; the other copy of the recombinase site was integrated at the EcoR I site at the "R4/R5" junction, ∼20 kb from the telomere with URA3 as the marker. Upon excision of the cassette, the TRP1 marker is left behind on the chromosome while URA3 is retained on the circle. For the experiment on long-term growth of cells with the excised circle, cells were plated on minimal medium lacking uracil after induction of recombination. Clones with the excised circle were identified by CHEF gel electrophoresis and by Southern blotting, probing for the expected junction fragment on the circle. To overcome poor growth arising from loss of the circle (which contains essential genes like RAD3), we integrated a centromere (CEN5) in the URA3 gene on the circle. Integrants were selected by growth on plates containing 5-fluoroorotic acid, which selects against expression of URA3 [J. D. Boeke, F. LaCroute, G. R. Fink, Mol. Gen. Genet. 197, 345 (1984)], and correct integration of the centromere fragment was confirmed by Southern blotting and hybridization.
    • (1984) Mol. Gen. Genet. , vol.197 , pp. 345
    • Boeke, J.D.1    LaCroute, F.2    Fink, G.R.3
  • 18
    • 0028568132 scopus 로고
    • Cells were embedded in agarose plugs and lysed [A. Gnirke, S. P. Iadonato, P. Y. Kwok, M. V. Olson, Genomics 24, 199 (1994)]. CHEF gel electrophoresis [G. Chu, D. Vollrath, R. W. Davis, Science 234, 1582 (1986)] was performed in a CHEF-DR II pulsed field electrophoresis system (Bio-Rad) for 19 hours at 200 V with a switch time of 23 s, followed by 19 hours at 200 V with a switch time of 46 s. The gel was 1% agarose in 0.5× tris-borate EDTA (TBE).
    • (1994) Genomics , vol.24 , pp. 199
    • Gnirke, A.1    Iadonato, S.P.2    Kwok, P.Y.3    Olson, M.V.4
  • 19
    • 0022965871 scopus 로고
    • Cells were embedded in agarose plugs and lysed [A. Gnirke, S. P. Iadonato, P. Y. Kwok, M. V. Olson, Genomics 24, 199 (1994)]. CHEF gel electrophoresis [G. Chu, D. Vollrath, R. W. Davis, Science 234, 1582 (1986)] was performed in a CHEF-DR II pulsed field electrophoresis system (Bio-Rad) for 19 hours at 200 V with a switch time of 23 s, followed by 19 hours at 200 V with a switch time of 46 s. The gel was 1% agarose in 0.5× tris-borate EDTA (TBE).
    • (1986) Science , vol.234 , pp. 1582
    • Chu, G.1    Vollrath, D.2    Davis, R.W.3
  • 22
    • 1842325374 scopus 로고    scopus 로고
    • note
    • 1 (Fig. 4), the culture was filtered, washed, and resuspended in isotopically normal minimal medium containing 1% sodium acetate, 0.5% ammonium sulfate, and α-factor, and split in half. Galactose (final concentration of 2%) was added to one portion; the other portion was the control, uninduced sample. After 5 hours, glucose was added to both portions (final concentration of 2%); 30 min thereafter, the cultures were shifted to 37°C and treated with Pronase, whereupon the cells progressed to the cdc7 block. The cells were released into S phase by shifting the culture to 23°C; samples were collected through S phase and processed for CsCl density gradient centrifugation (5). For excision at the nocodazole block, cells grown in isotopically dense medium were synchronized with α-factor as before, and nocadazole was added to the culture (final concentration of 10 μg/ ml) before release from the α-pheromone block. Once the culture had arrested as predominantly large-budded cells, it was split in two and galactose was added to one portion. After 5 hours, both portions were filtered, washed, and resuspended in isotopically normal minimal medium containing 2% glucose and incubated at 37°C until the cells reached the cdc7 block. The cells were then released into S phase and samples were collected as before. Small samples were also collected at various steps through the whole procedure and analyzed by flow cytometry to confirm the cell-cycle arrests (16).
  • 24
    • 0026845839 scopus 로고
    • O. M. Aparicio, B. L. Billington, D. E. Gottschling, Cell 66, 1279 (1991); D. H. Rivier and J. Rine, Curr. Opin. Genet. Dev. 2, 286 (1992).
    • (1992) Curr. Opin. Genet. Dev. , vol.2 , pp. 286
    • Rivier, D.H.1    Rine, J.2
  • 28
    • 0027423154 scopus 로고
    • F. Palladino et al., Cell 75, 543 (1993).
    • (1993) Cell , vol.75 , pp. 543
    • Palladino, F.1
  • 30
    • 0026550902 scopus 로고
    • J. F. Diffley and J. H. Cocker, Nature 357, 169 (1992); S. P. Bell and B. Stillman, ibid., p. 128.
    • (1992) Nature , vol.357 , pp. 169
    • Diffley, J.F.1    Cocker, J.H.2
  • 31
    • 0026607331 scopus 로고    scopus 로고
    • J. F. Diffley and J. H. Cocker, Nature 357, 169 (1992); S. P. Bell and B. Stillman, ibid., p. 128.
    • Nature , pp. 128
    • Bell, S.P.1    Stillman, B.2
  • 34
    • 0027367572 scopus 로고
    • K. M. Hennessy, C. D. Clark, D. Botstein, Genes Dev. 4, 2252 (1990); H. Van, A. M. Merchant, B. K. Tye, ibid. 7, 2149 (1993).
    • (1993) Genes Dev. , vol.7 , pp. 2149
    • Van, H.1    Merchant, A.M.2    Tye, B.K.3
  • 36
    • 1842286871 scopus 로고    scopus 로고
    • note
    • We are grateful to M. Gartenberg and K. Kolor for gifts of plasmids and strains and to M. Hoang for technical assistance. We also thank A. Donaldson, K. Kolor, and A. van Brabant for their helpful comments on the manuscript. Supported by National Institutes of General Medical Sciences grant 18926 to B.J.B. and W.L.F.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.