-
2
-
-
0026511056
-
-
D. E. Levin and E. Bartlett-Heubusch, J. Cell Biol. 116, 1221 (1992); G. Paravicini et al., Mol. Cell. Biol. 12, 4896 (1992); D. E. Levin and B. Errede, Curr. Opin. Cell Biol. 7, 197 (1995).
-
(1992)
J. Cell Biol.
, vol.116
, pp. 1221
-
-
Levin, D.E.1
Bartlett-Heubusch, E.2
-
3
-
-
0026693868
-
-
D. E. Levin and E. Bartlett-Heubusch, J. Cell Biol. 116, 1221 (1992); G. Paravicini et al., Mol. Cell. Biol. 12, 4896 (1992); D. E. Levin and B. Errede, Curr. Opin. Cell Biol. 7, 197 (1995).
-
(1992)
Mol. Cell. Biol.
, vol.12
, pp. 4896
-
-
Paravicini, G.1
-
4
-
-
0028920365
-
-
D. E. Levin and E. Bartlett-Heubusch, J. Cell Biol. 116, 1221 (1992); G. Paravicini et al., Mol. Cell. Biol. 12, 4896 (1992); D. E. Levin and B. Errede, Curr. Opin. Cell Biol. 7, 197 (1995).
-
(1995)
Curr. Opin. Cell Biol.
, vol.7
, pp. 197
-
-
Levin, D.E.1
Errede, B.2
-
5
-
-
0026584373
-
-
K. Lee and D. Levin, Mol. Cell. Biol. 12, 172 (1992); K. Irie et al., ibid. 13, 3076 (1993); K. S. Lee et al., ibid., p. 3067.
-
(1992)
Mol. Cell. Biol.
, vol.12
, pp. 172
-
-
Lee, K.1
Levin, D.2
-
6
-
-
0027192944
-
-
K. Lee and D. Levin, Mol. Cell. Biol. 12, 172 (1992); K. Irie et al., ibid. 13, 3076 (1993); K. S. Lee et al., ibid., p. 3067.
-
(1993)
Mol. Cell. Biol.
, vol.13
, pp. 3076
-
-
Irie, K.1
-
7
-
-
0026584373
-
-
K. Lee and D. Levin, Mol. Cell. Biol. 12, 172 (1992); K. Irie et al., ibid. 13, 3076 (1993); K. S. Lee et al., ibid., p. 3067.
-
Mol. Cell. Biol.
, pp. 3067
-
-
Lee, K.S.1
-
9
-
-
0027761005
-
-
C. Mazzoni, P. Zarzov, A. Rambourg, C. Mann, J. Cell Biol. 123, 1821 (1993).
-
(1993)
J. Cell Biol.
, vol.123
, pp. 1821
-
-
Mazzoni, C.1
Zarzov, P.2
Rambourg, A.3
Mann, C.4
-
10
-
-
0028952804
-
-
C. J. DiComo, H. Chang, K. T. Arndt, Mol. Cell. Biol. 15, 1835 (1995); B. A. Morgan, N. Bouquin, G. F. Merrill, L. H. Johnston, EMBO J. 14, 5679 (1995); N. J. Marini et al., ibid. 15, 3040 (1996).
-
(1995)
Mol. Cell. Biol.
, vol.15
, pp. 1835
-
-
DiComo, C.J.1
Chang, H.2
Arndt, K.T.3
-
11
-
-
0028805253
-
-
C. J. DiComo, H. Chang, K. T. Arndt, Mol. Cell. Biol. 15, 1835 (1995); B. A. Morgan, N. Bouquin, G. F. Merrill, L. H. Johnston, EMBO J. 14, 5679 (1995); N. J. Marini et al., ibid. 15, 3040 (1996).
-
(1995)
EMBO J.
, vol.14
, pp. 5679
-
-
Morgan, B.A.1
Bouquin, N.2
Merrill, G.F.3
Johnston, L.H.4
-
12
-
-
0030007679
-
-
C. J. DiComo, H. Chang, K. T. Arndt, Mol. Cell. Biol. 15, 1835 (1995); B. A. Morgan, N. Bouquin, G. F. Merrill, L. H. Johnston, EMBO J. 14, 5679 (1995); N. J. Marini et al., ibid. 15, 3040 (1996).
-
(1996)
EMBO J.
, vol.15
, pp. 3040
-
-
Marini, N.J.1
-
16
-
-
0024550203
-
-
B. J. Andrews and I. Herskowitz, Cell 57, 21 (1989); M. Primig, S. Sockanathan, H. Auer, K. Nasmyth, Nature 358, 593 (1992); J. Sidorova and L. Breeden, Mol. Cell. Biol. 13, 1069 (1993).
-
(1989)
Cell
, vol.57
, pp. 21
-
-
Andrews, B.J.1
Herskowitz, I.2
-
17
-
-
0026649405
-
-
B. J. Andrews and I. Herskowitz, Cell 57, 21 (1989); M. Primig, S. Sockanathan, H. Auer, K. Nasmyth, Nature 358, 593 (1992); J. Sidorova and L. Breeden, Mol. Cell. Biol. 13, 1069 (1993).
-
(1992)
Nature
, vol.358
, pp. 593
-
-
Primig, M.1
Sockanathan, S.2
Auer, H.3
Nasmyth, K.4
-
18
-
-
0027500140
-
-
B. J. Andrews and I. Herskowitz, Cell 57, 21 (1989); M. Primig, S. Sockanathan, H. Auer, K. Nasmyth, Nature 358, 593 (1992); J. Sidorova and L. Breeden, Mol. Cell. Biol. 13, 1069 (1993).
-
(1993)
Mol. Cell. Biol.
, vol.13
, pp. 1069
-
-
Sidorova, J.1
Breeden, L.2
-
20
-
-
84967769748
-
-
K. Nasmyth and L. Dirick, ibid., p. 995; M. J. Fernandez-Sarabia, A. Sutton, T. Zhong, K. T. Arndt, Genes Dev. 6, 2417 (1992); M. Tyers, G. Tokiwa, B. Futcher, EMBO J. 12, 1955 (1993).
-
Cell
, pp. 995
-
-
Nasmyth, K.1
Dirick, L.2
-
21
-
-
0027102212
-
-
K. Nasmyth and L. Dirick, ibid., p. 995; M. J. Fernandez-Sarabia, A. Sutton, T. Zhong, K. T. Arndt, Genes Dev. 6, 2417 (1992); M. Tyers, G. Tokiwa, B. Futcher, EMBO J. 12, 1955 (1993).
-
(1992)
Genes Dev.
, vol.6
, pp. 2417
-
-
Fernandez-Sarabia, M.J.1
Sutton, A.2
Zhong, T.3
Arndt, K.T.4
-
22
-
-
0027173154
-
-
K. Nasmyth and L. Dirick, ibid., p. 995; M. J. Fernandez-Sarabia, A. Sutton, T. Zhong, K. T. Arndt, Genes Dev. 6, 2417 (1992); M. Tyers, G. Tokiwa, B. Futcher, EMBO J. 12, 1955 (1993).
-
(1993)
EMBO J.
, vol.12
, pp. 1955
-
-
Tyers, M.1
Tokiwa, G.2
Futcher, B.3
-
23
-
-
0028559010
-
-
F. H. Espinoza, J. Ogas, I. Herskowitz, D. O. Morgan, Science 266, 1388 (1994); V. Measday, L. Moore, J. Ogas, M. Tyers, B. Andrews, ibid., p. 1391.
-
(1994)
Science
, vol.266
, pp. 1388
-
-
Espinoza, F.H.1
Ogas, J.2
Herskowitz, I.3
Morgan, D.O.4
-
24
-
-
0028589197
-
-
F. H. Espinoza, J. Ogas, I. Herskowitz, D. O. Morgan, Science 266, 1388 (1994); V. Measday, L. Moore, J. Ogas, M. Tyers, B. Andrews, ibid., p. 1391.
-
Science
, pp. 1391
-
-
Measday, V.1
Moore, L.2
Ogas, J.3
Tyers, M.4
Andrews, B.5
-
25
-
-
0024834127
-
-
H. E. Richardson, C. Wittenberg, F. Cross, S. I. Reed, Cell 59, 1127 (1989); F. R. Cross, Mol. Cell. Biol. 10, 6482 (1990).
-
(1989)
Cell
, vol.59
, pp. 1127
-
-
Richardson, H.E.1
Wittenberg, C.2
Cross, F.3
Reed, S.I.4
-
26
-
-
0025200729
-
-
H. E. Richardson, C. Wittenberg, F. Cross, S. I. Reed, Cell 59, 1127 (1989); F. R. Cross, Mol. Cell. Biol. 10, 6482 (1990).
-
(1990)
Mol. Cell. Biol.
, vol.10
, pp. 6482
-
-
Cross, F.R.1
-
28
-
-
0026600968
-
-
C. Costigan, S. Gehrung, M. Snyder, Mol. Cell. Biol. 12, 1162 (1992); C. Costigan and M. Snyder, ibid. 14, 2391 (1994).
-
(1992)
Mol. Cell. Biol.
, vol.12
, pp. 1162
-
-
Costigan, C.1
Gehrung, S.2
Snyder, M.3
-
29
-
-
0028331458
-
-
C. Costigan, S. Gehrung, M. Snyder, Mol. Cell. Biol. 12, 1162 (1992); C. Costigan and M. Snyder, ibid. 14, 2391 (1994).
-
(1994)
Mol. Cell. Biol.
, vol.14
, pp. 2391
-
-
Costigan, C.1
Snyder, M.2
-
32
-
-
0024539080
-
-
M. Snyder, J. Cell Biol. 108, 1419 (1989); S. Gehrung and M. Snyder, ibid. 111, 1451 (1990).
-
(1989)
J. Cell Biol.
, vol.108
, pp. 1419
-
-
Snyder, M.1
-
34
-
-
0027527033
-
-
A. Amon, M. Tyers, B. Futcher, K. Nasmyth, Cell 74, 993 (1993); J. M. Sidorova, G. E. Mikesell, L. L. Breeden, Mol. Biol. Cell 6, 1641 (1995).
-
(1993)
Cell
, vol.74
, pp. 993
-
-
Amon, A.1
Tyers, M.2
Futcher, B.3
Nasmyth, K.4
-
35
-
-
0028820406
-
-
A. Amon, M. Tyers, B. Futcher, K. Nasmyth, Cell 74, 993 (1993); J. M. Sidorova, G. E. Mikesell, L. L. Breeden, Mol. Biol. Cell 6, 1641 (1995).
-
(1995)
Mol. Biol. Cell
, vol.6
, pp. 1641
-
-
Sidorova, J.M.1
Mikesell, G.E.2
Breeden, L.L.3
-
36
-
-
0029935418
-
-
R. Treisman, Curr. Opin. Cell Biol. 8, 205 (1996); J. E. Slansky and P. J. Farnham, Curr. Top. Microbiol. Immunol. 208, 1 (1996).
-
(1996)
Curr. Opin. Cell Biol.
, vol.8
, pp. 205
-
-
Treisman, R.1
-
38
-
-
1842414468
-
-
note
-
Congenic wild-type (WT) (Y762), bck1Δ::TRP1 (Y782), slt2Δ::LEU2 (Y783), and swi4Δ::HIS3 (YK74) strains were prepared (11, 16). High-copy plasmids containing the yeast 2μ origin of replication and either SLT2 (16), PCL1 (11), PCL2 (pBA623; Hind III-Sal I fragment from a genomic library plasmid cloned into pRS426), PHO85 (BK55; YEp24 genomic library plasmid), SWI4 (B327; 8-kb Sph I fragment including SWI4 subcloned into YEp352), or MBP1 [BK72; polymerase chain reaction product including MBP1 and flanking sequence (shown to be functional) cloned into YEp24] were transformed into slt2Δ and bck1Δ strains. These plasmids are expected to be present at ∼20 copies per cell (33). Transformants were streaked on plates containing synthetic media lacking uracil (34) and incubated for 3 days at 37°C.
-
-
-
-
40
-
-
1842300069
-
-
note
-
slt2Δ cells containing either YEp24, or SWI4, PCL1, or PCL2 high-copy plasmids (Fig. 1) were grown to late-log phase in 5-ml cultures of synthetic medium lacking uracil (with or without 1 M sorbitol); cells were diluted to an absorbance at 600 nm of 0.1 and grown for 3 hours in fresh synthetic complete medium. Cells were then centrifuged, resuspended, and incubated for 2 hours in 5 ml of rich medium with or without 1 M sorbitol. α-Factor (Sigma) was then added (5 μg/ml), and after 1 hour an equivalent dose of pheromone was added to the culture. The incubation was continued for 1 hour, and cells were fixed with 3.7% formaldehyde and washed three times with phosphate-buffered saline. Cells were observed by differential-interference microscopy, and the level of suppression of slt2Δ mutant phenotypes was quantified. Cells were either lysed, budded, or arrested in response to pheromone. Cells arrested in response to mating factor were divided into two classes on the basis of projection length: either ≥ or < one-third the diameter of the cell at the widest point. "Other" category includes cells with more than one projection or cells that have undergone polarized growth events that could not clearly be classified as buds or mating projections.
-
-
-
-
41
-
-
0027191189
-
-
M. Peter, A. Gartner, J. Horecker, G. Ammerer, Cell 73, 747 (1993); M. Tyers and B. Futcher, Mol. Cell. Biol. 13, 5659 (1993).
-
(1993)
Cell
, vol.73
, pp. 747
-
-
Peter, M.1
Gartner, A.2
Horecker, J.3
Ammerer, G.4
-
42
-
-
0027261380
-
-
M. Peter, A. Gartner, J. Horecker, G. Ammerer, Cell 73, 747 (1993); M. Tyers and B. Futcher, Mol. Cell. Biol. 13, 5659 (1993).
-
(1993)
Mol. Cell. Biol.
, vol.13
, pp. 5659
-
-
Tyers, M.1
Futcher, B.2
-
43
-
-
0031036674
-
-
Y. Ho, S. Mason, R. Kobayashi, M. Hoekstra, B. Andrews, Proc. Natl. Acad. Sci. U.S.A. 94, 581 (1997).
-
(1997)
Proc. Natl. Acad. Sci. U.S.A.
, vol.94
, pp. 581
-
-
Ho, Y.1
Mason, S.2
Kobayashi, R.3
Hoekstra, M.4
Andrews, B.5
-
44
-
-
1842368708
-
-
note
-
Affinity-purified anti-Swi6 was used to immunoprecipitate Swi6p from 1 mg of total protein isolated from cells containing high-copy SLT2:HA; the cells had been incubated at 39°C for 1 hour. Immunoprecipitates were resuspended in 45 μl of NEB (New England Biolabs) Buffer 3 with 1% SDS. After 5 min of incubation at 37°C, the supernatant was removed and diluted to 0.1% SDS in NEB Buffer 3 with either 30 U of calf intestinal alkaline phosphatase (CIP) or 30 U of CIP together with 4 mM sodium molybdate (CIP + inhib.). Phosphatase reactions were incubated for 30 min at 37°C, and then Swi6p was reimmunoprecipitated from the phosphatase reaction.
-
-
-
-
45
-
-
11944273348
-
-
Y. Kamada, U. S. Jung, J. Piotrowski, D. E. Levin, Genes Dev. 9, 1559 (1995).
-
(1995)
Genes Dev.
, vol.9
, pp. 1559
-
-
Kamada, Y.1
Jung, U.S.2
Piotrowski, J.3
Levin, D.E.4
-
46
-
-
1842289428
-
-
note
-
32P]ATP, and the reaction was allowed to proceed for 50 min. Half of each reaction was separated in a 10% polyacrylamide gel and blotted; the filter was used for autoradiography.
-
-
-
-
49
-
-
1842327030
-
-
E. Rosengurt, A. Rodriguez-Pena, M. Coombs, J. Sinnet-Smith, Proc. Natl. Acad. Sci. U.S.A. 81, 5748 (1984); K. Kaibuchi, Y. Takai, Y. Nishizuka, J. Biol. Chem. 260, 1366 (1985); D. A. Persons, W. O. Wilkison, R. M. Bell, O. J. Finn, Cell 52, 447 (1988); H. Michak et al., J. Biol. Chem. 268, 110 (1993).
-
(1984)
Proc. Natl. Acad. Sci. U.S.A.
, vol.81
, pp. 5748
-
-
Rosengurt, E.1
Rodriguez-Pena, A.2
Coombs, M.3
Sinnet-Smith, J.4
-
50
-
-
0021913289
-
-
E. Rosengurt, A. Rodriguez-Pena, M. Coombs, J. Sinnet-Smith, Proc. Natl. Acad. Sci. U.S.A. 81, 5748 (1984); K. Kaibuchi, Y. Takai, Y. Nishizuka, J. Biol. Chem. 260, 1366 (1985); D. A. Persons, W. O. Wilkison, R. M. Bell, O. J. Finn, Cell 52, 447 (1988); H. Michak et al., J. Biol. Chem. 268, 110 (1993).
-
(1985)
J. Biol. Chem.
, vol.260
, pp. 1366
-
-
Kaibuchi, K.1
Takai, Y.2
Nishizuka, Y.3
-
51
-
-
0024284812
-
-
E. Rosengurt, A. Rodriguez-Pena, M. Coombs, J. Sinnet-Smith, Proc. Natl. Acad. Sci. U.S.A. 81, 5748 (1984); K. Kaibuchi, Y. Takai, Y. Nishizuka, J. Biol. Chem. 260, 1366 (1985); D. A. Persons, W. O. Wilkison, R. M. Bell, O. J. Finn, Cell 52, 447 (1988); H. Michak et al., J. Biol. Chem. 268, 110 (1993).
-
(1988)
Cell
, vol.52
, pp. 447
-
-
Persons, D.A.1
Wilkison, W.O.2
Bell, R.M.3
Finn, O.J.4
-
52
-
-
1842412131
-
-
E. Rosengurt, A. Rodriguez-Pena, M. Coombs, J. Sinnet-Smith, Proc. Natl. Acad. Sci. U.S.A. 81, 5748 (1984); K. Kaibuchi, Y. Takai, Y. Nishizuka, J. Biol. Chem. 260, 1366 (1985); D. A. Persons, W. O. Wilkison, R. M. Bell, O. J. Finn, Cell 52, 447 (1988); H. Michak et al., J. Biol. Chem. 268, 110 (1993).
-
(1993)
J. Biol. Chem.
, vol.268
, pp. 110
-
-
Michak, H.1
-
54
-
-
77957772100
-
-
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
-
F. Sherman, G. Fink, J. Hicks, Cold Spring Harbor Laboratories - Methods in Yeast Genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1986).
-
(1986)
Cold Spring Harbor Laboratories - Methods in Yeast Genetics
-
-
Sherman, F.1
Fink, G.2
Hicks, J.3
-
55
-
-
0003448569
-
-
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
-
E. Harlow and D. Lane, Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988).
-
(1988)
Antibodies: A Laboratory Manual
-
-
Harlow, E.1
Lane, D.2
-
56
-
-
1842332920
-
-
note
-
Proteins were separated by SDS-polyacrylamide gel electrophoresis (PAGE; 10% gel), blotted onto Immobilon-P (Millipore), probed with affinity-purified anti-Swi6p, and primary antibodies were detected with alkaline phosphatase-conjugated goat anti-rabbit IgG and the bromochloroindolyl phosphate-nitro blue tetrazolium (BCIP-NBT) color substrate reaction (35).
-
-
-
-
58
-
-
1842332323
-
-
note
-
Samples for two-dimensional electrophoresis were suspended in SDS sample buffer, then treated so that the first-dimension sample buffer contained 9.5 M urea, 2.0% Triton X-100, 5% β-mercaptoethanol, and 1.6% Bio-Lyte 5/7 and 0.4% Bio-Lyte 3/10 ampholytes (Bio-Rad). First-dimension isoelectric focusing and second-dimension SDS-PAGE (10% gel) were performed as recommended (Mini-PROTEAN II, Bio-Rad). The SDS-PAGE gel was blotted (36) and exposed for autoradiography (29).
-
-
-
-
59
-
-
1842405006
-
-
note
-
We thank C. Mann for strains and plasmids, Y. Ho for providing antibodies and extracts, and Y. Barral, C. Costigan, and C. Crews for critical comments on the manuscript. Supported by grants from the National Institutes of Health (GM36494) (M.S.), the Medical Research Council of Canada (B.A.), a Howard Hughes Medical Institute predoctoral fellowship (K.M.), and a Connaught predoctoral scholarship (K.B.).
-
-
-
|