-
2
-
-
0026452949
-
-
L Hartwell, Cell 71, 543 (1992), T A. Weinert and D. Lydall, Semin Cancer Biol. 4, 129 (1993).
-
(1992)
Cell
, vol.71
, pp. 543
-
-
Hartwell, L.1
-
6
-
-
0026596980
-
-
R Rowley, S Subramani, P. G. Young, ibid , p 1335, T. Enoch, A. M. Carr, P. Nurse, Genes Dev. 6, 2035 (1993).
-
EMBO J.
, pp. 1335
-
-
Rowley, R.1
Subramani, S.2
Young, P.G.3
-
7
-
-
0026482601
-
-
R Rowley, S Subramani, P. G. Young, ibid , p 1335, T. Enoch, A. M. Carr, P. Nurse, Genes Dev. 6, 2035 (1993).
-
(1993)
Genes Dev.
, vol.6
, pp. 2035
-
-
Enoch, T.1
Carr, A.M.2
Nurse, P.3
-
10
-
-
0027212282
-
-
T. A. Weinert and L H. Hartwell, Genetics 134, 63 (1993); T. A Weinert, G L. Kiser, L. H. Hartwell, Genes Dev. 8, 652 (1994).
-
(1993)
Genetics
, vol.134
, pp. 63
-
-
Weinert, T.A.1
Hartwell, L.H.2
-
11
-
-
0028353634
-
-
T. A. Weinert and L H. Hartwell, Genetics 134, 63 (1993); T. A Weinert, G L. Kiser, L. H. Hartwell, Genes Dev. 8, 652 (1994).
-
(1994)
Genes Dev.
, vol.8
, pp. 652
-
-
Weinert, T.A.1
Kiser, G.L.2
Hartwell, L.H.3
-
12
-
-
3142647895
-
-
P. Nurse and Y Bissett, Nature 292, 448 (1981), P. Nurse, ibid. 344, 503 (1990).
-
(1981)
Nature
, vol.292
, pp. 448
-
-
Nurse, P.1
Bissett, Y.2
-
13
-
-
0025246110
-
-
P. Nurse and Y Bissett, Nature 292, 448 (1981), P. Nurse, ibid. 344, 503 (1990).
-
(1990)
Nature
, vol.344
, pp. 503
-
-
Nurse, P.1
-
14
-
-
0026509543
-
-
M Tyers, G. Tokiwa, R. Nash, B Futcher, EMBO J. 11, 1773 (1992)
-
(1992)
EMBO J.
, vol.11
, pp. 1773
-
-
Tyers, M.1
Tokiwa, G.2
Nash, R.3
Futcher, B.4
-
15
-
-
4243061361
-
-
+ coding sequence to generate the chk1:ep allele
-
+ coding sequence to generate the chk1:ep allele
-
-
-
-
16
-
-
4243072630
-
-
chk1 ep was detected in the cdc17-K42 mutant at the permissive temperature, suggesting that because of the low activity of DNA ligase, the DNA damage checkpoint is constitutively required to prevent cells from entering mitosis with unligated DNA
-
(1977)
Cell
, vol.12
, pp. 101
-
-
Nasmyth, K.A.1
-
17
-
-
4243059149
-
-
chk1 ep was detected in lysates of these cells
-
chk1 ep was detected in lysates of these cells
-
-
-
-
19
-
-
4243093630
-
-
Septation index (SI) was measured by fixing cells in 70% ethanol, washing with water, and then incubating with calcofluor to stain the septa. An asynchronous population of logarithmically growing S. pombe has an SI of ∼15% By 2 to 3 hours after incubation with 12 mM HU, the SI had decreased to ≤2%, indicating that the cells were no longer traversing the cell cycle
-
Septation index (SI) was measured by fixing cells in 70% ethanol, washing with water, and then incubating with calcofluor to stain the septa. An asynchronous population of logarithmically growing S. pombe has an SI of ∼15% By 2 to 3 hours after incubation with 12 mM HU, the SI had decreased to ≤2%, indicating that the cells were no longer traversing the cell cycle.
-
-
-
-
20
-
-
0342436767
-
-
- [T A Kunkel, J D Roberts, R A Zakour, Methods Enzymol. 155, 166 (1987)]. The oligonucleotide used was 5′-TATGCTGTCGCATTTGTCAAT-3′. in which the underlined bases are altered from the wild-type sequence. A small fragment encompassing the mutated site was cloned into pBluescript (Stratagene) and completely sequenced before cloning back into the cDNA expression vector
-
(1987)
Methods Enzymol.
, vol.155
, pp. 166
-
-
Kunkel, T.A.1
Roberts, J.D.2
Zakour, R.A.3
-
22
-
-
4243065924
-
-
The cDNAs were cloned into pREP81, in which expression is controlled from a weakened nmt1 promoter [G. Basi, E Schmid, K Maundrell, Gene 114, 59 (1993)]. The nmt1 promoter is repressive by thiamine [K. Maundrell, J Biol Chem. 265, 10857 (1990)].
-
(1993)
Gene
, vol.114
, pp. 59
-
-
Basi, G.1
Schmid, E.2
Maundrell, K.3
-
23
-
-
0025298571
-
-
The cDNAs were cloned into pREP81, in which expression is controlled from a weakened nmt1 promoter [G. Basi, E Schmid, K Maundrell, Gene 114, 59 (1993)]. The nmt1 promoter is repressive by thiamine [K. Maundrell, J Biol Chem. 265, 10857 (1990)].
-
(1990)
J Biol Chem.
, vol.265
, pp. 10857
-
-
Maundrell, K.1
-
24
-
-
4243182243
-
-
chk1 ep expressed from the genomic locus
-
chk1 ep expressed from the genomic locus
-
-
-
-
25
-
-
0028101138
-
-
J. C Ford et al., Science 265, 533 (1994).
-
(1994)
Science
, vol.265
, pp. 533
-
-
Ford, J.C.1
-
26
-
-
4243109218
-
-
unpublished data
-
N. C. Walworth, unpublished data.
-
-
-
Walworth, N.C.1
-
28
-
-
4243150357
-
-
2] containing protoase inhibitors that is compatible with the activity of lambda protein phosphatase (New England Biolabs) Lysate (25 μg) was incubated with 40 U of phosphatase in a volume of 30 μl for 30 min at 3O°C Control reactions were incubated on ice. Reactions were terminated by addition of 30 μl of 2 × Laemmli sample buffer and heating to 95°C for 5 min.
-
2] containing protoase inhibitors that is compatible with the activity of lambda protein phosphatase (New England Biolabs) Lysate (25 μg) was incubated with 40 U of phosphatase in a volume of 30 μl for 30 min at 3O°C Control reactions were incubated on ice. Reactions were terminated by addition of 30 μl of 2 × Laemmli sample buffer and heating to 95°C for 5 min.
-
-
-
-
29
-
-
0025784689
-
-
J M. Murray, A M Carr, A. R Lehmann, F. Z. Watts, Nucleic Acids Res. 19, 3525 (1991); P Sunnerhagen, B. Seaton, A. Nasim, S Subramani, Mol. Cell Biol 10, 3750 (1990), A. M. Carr, personal communication
-
(1991)
Nucleic Acids Res.
, vol.19
, pp. 3525
-
-
Murray, J.M.1
Carr, A.M.2
Lehmann, A.R.3
Watts, F.Z.4
-
30
-
-
0025351487
-
-
J M. Murray, A M Carr, A. R Lehmann, F. Z. Watts, Nucleic Acids Res. 19, 3525 (1991); P Sunnerhagen, B. Seaton, A. Nasim, S Subramani, Mol. Cell Biol 10, 3750 (1990), A. M. Carr, personal communication
-
(1990)
Mol. Cell Biol
, vol.10
, pp. 3750
-
-
Sunnerhagen, P.1
Seaton, B.2
Nasim, A.3
Subramani, S.4
-
31
-
-
0025784689
-
-
personal communication
-
J M. Murray, A M Carr, A. R Lehmann, F. Z. Watts, Nucleic Acids Res. 19, 3525 (1991); P Sunnerhagen, B. Seaton, A. Nasim, S Subramani, Mol. Cell Biol 10, 3750 (1990), A. M. Carr, personal communication
-
-
-
Carr, A.M.1
-
34
-
-
0029057336
-
-
K. Savitsky et al., Science 268, 1749 (1995).
-
(1995)
Science
, vol.268
, pp. 1749
-
-
Savitsky, K.1
-
35
-
-
0025780420
-
-
R. A. Gatti et al., Medicine 70, 99 (1991).
-
(1991)
Medicine
, vol.70
, pp. 99
-
-
Gatti, R.A.1
-
36
-
-
0023244806
-
-
M Swift et al., N. Engl. J. Med. 316, 1289 (1987), A. L Borreson et al., Cancer 2, 339 (1990); M. Swift et al., N Engl. J. Med 325, 1831 (1991).
-
(1987)
N. Engl. J. Med.
, vol.316
, pp. 1289
-
-
Swift, M.1
-
37
-
-
0025683343
-
-
M Swift et al., N. Engl. J. Med. 316, 1289 (1987), A. L Borreson et al., Cancer 2, 339 (1990); M. Swift et al., N Engl. J. Med 325, 1831 (1991).
-
(1990)
Cancer
, vol.2
, pp. 339
-
-
Borreson, A.L.1
-
38
-
-
0026409331
-
-
M Swift et al., N. Engl. J. Med. 316, 1289 (1987), A. L Borreson et al., Cancer 2, 339 (1990); M. Swift et al., N Engl. J. Med 325, 1831 (1991).
-
(1991)
N Engl. J. Med
, vol.325
, pp. 1831
-
-
Swift, M.1
-
39
-
-
0026686944
-
-
B L. Seaton et al , Gene 119, 83 (1992); G. Jimenez et al., Proc. Natl. Acad Sci. U.S.A 89, 4952 (1992). The published sequence of rad3 is incomplete, and the missing domains include the lipid kinase catalytic domain (A. M. Carr and N Bentley. personal communication).
-
(1992)
Gene
, vol.119
, pp. 83
-
-
Seaton, B.L.1
-
40
-
-
0026636130
-
-
B L. Seaton et al , Gene 119, 83 (1992); G. Jimenez et al., Proc. Natl. Acad Sci. U.S.A 89, 4952 (1992). The published sequence of rad3 is incomplete, and the missing domains include the lipid kinase catalytic domain (A. M. Carr and N Bentley. personal communication).
-
(1992)
Proc. Natl. Acad Sci. U.S.A
, vol.89
, pp. 4952
-
-
Jimenez, G.1
-
41
-
-
0026686944
-
-
personal communication
-
B L. Seaton et al , Gene 119, 83 (1992); G. Jimenez et al., Proc. Natl. Acad Sci. U.S.A 89, 4952 (1992). The published sequence of rad3 is incomplete, and the missing domains include the lipid kinase catalytic domain (A. M. Carr and N Bentley. personal communication).
-
-
-
Carr, A.M.1
Bentley, N.2
-
42
-
-
0019506288
-
-
F. Zampetti-Bosseler and D Scott, Int. J Radiat. Biol 39, 547 (1981); M. D. Ford, L. Martin, M. F. Lavin, Mutat. Res. 125, 115 (1984), H Beamish, K K. Khanna, M. F. Lavm, Radiat. Res. 138, S130 (1994).
-
(1981)
Int. J Radiat. Biol
, vol.39
, pp. 547
-
-
Zampetti-Bosseler, F.1
Scott, D.2
-
43
-
-
0021366742
-
-
F. Zampetti-Bosseler and D Scott, Int. J Radiat. Biol 39, 547 (1981); M. D. Ford, L. Martin, M. F. Lavin, Mutat. Res. 125, 115 (1984), H Beamish, K K. Khanna, M. F. Lavm, Radiat. Res. 138, S130 (1994).
-
(1984)
Mutat. Res.
, vol.125
, pp. 115
-
-
Ford, M.D.1
Martin, L.2
Lavin, M.F.3
-
44
-
-
0028269782
-
-
F. Zampetti-Bosseler and D Scott, Int. J Radiat. Biol 39, 547 (1981); M. D. Ford, L. Martin, M. F. Lavin, Mutat. Res. 125, 115 (1984), H Beamish, K K. Khanna, M. F. Lavm, Radiat. Res. 138, S130 (1994).
-
(1994)
Radiat. Res.
, vol.138
-
-
Beamish, H.1
Khanna, K.K.2
Lavm, M.F.3
-
46
-
-
4243154809
-
-
3 and 50 mM NaF, and then lysing in the same solution containing protease inhibitors by vortexing with glass beads. After recovery from the beads, the lysate was centrifuged briefly at 5000g to remove unbroken cells For immunoblot analysis, 15 to 25 μg of cell extract was separated on an 8% SDS-polyacrylarnide gel and transferred to nitrocellulose. Blocking of nonspecific sites, antibody incubations, and all washes were performed in PBS containing 1% dried nonfat milk and 0.05% Tween-20. The 12CA5 antibody directed against the HA epitopes was used at a 1:1000 dilution, and the secondary, horseradish peroxidase - coupled goat antibodies to mouse immunoglobulm G (TAGO), at a 1.10,000 dilution. After a final rinse with PBS containing 0 05% Tween-20, immune complexes were detected with the ECL system (Amersham)
-
3 and 50 mM NaF, and then lysing in the same solution containing protease inhibitors by vortexing with glass beads. After recovery from the beads, the lysate was centrifuged briefly at 5000g to remove unbroken cells For immunoblot analysis, 15 to 25 μg of cell extract was separated on an 8% SDS-polyacrylarnide gel and transferred to nitrocellulose. Blocking of nonspecific sites, antibody incubations, and all washes were performed in PBS containing 1% dried nonfat milk and 0.05% Tween-20. The 12CA5 antibody directed against the HA epitopes was used at a 1:1000 dilution, and the secondary, horseradish peroxidase - coupled goat antibodies to mouse immunoglobulm G (TAGO), at a 1.10,000 dilution. After a final rinse with PBS containing 0 05% Tween-20, immune complexes were detected with the ECL system (Amersham)
-
-
-
-
47
-
-
4243193344
-
-
We thank members of the Bernards laboratory as well as M Smeets and A. Begg at the Netherlands Cancer Institute for helpful discussions, A. M. Carr for providing strains containing the radΔ alleles, and B Nefsky for critical reading of the manuscript Supported by grants to R B. from the Netherlands Organization for Scientific Research (NWO) and the NIH, and from start-up funds provided to N.W. from UMDNJ-Robert Wood Johnson Medical School
-
We thank members of the Bernards laboratory as well as M Smeets and A. Begg at the Netherlands Cancer Institute for helpful discussions, A. M. Carr for providing strains containing the radΔ alleles, and B Nefsky for critical reading of the manuscript Supported by grants to R B. from the Netherlands Organization for Scientific Research (NWO) and the NIH, and from start-up funds provided to N.W. from UMDNJ-Robert Wood Johnson Medical School
-
-
-
|