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Volumn 284, Issue 5416, 1999, Pages 977-980

Roles of phosphorylation sites in regulating activity of the transcription factor pho4

Author keywords

[No Author keywords available]

Indexed keywords

FUNGAL PROTEIN; TRANSCRIPTION FACTOR;

EID: 0033532281     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.284.5416.977     Document Type: Article
Times cited : (261)

References (35)
  • 8
    • 0028894785 scopus 로고
    • K. R. Schneider, R. L. Smith, E. K. O'Shea, ibid. 266, 122 (1994); N. Ogawa et al., Mol. Cell. Biol. 15, 997 (1995).
    • (1995) Mol. Cell. Biol. , vol.15 , pp. 997
    • Ogawa, N.1
  • 10
    • 0344277932 scopus 로고    scopus 로고
    • note
    • Previous studies demonstrate that Pho4 mutants containing Ser→Ala substitutions of a subset of the phosphorylation sites can be efficiently phosphorylated in vitro (6).
  • 11
    • 0344709944 scopus 로고    scopus 로고
    • note
    • SA1465D23-GFP, containing Ser→Ala substitutions at sites 1, 4, and 6, and Ser→Asp substitutions at sites 2 and 3, is not exported from the nucleus (8).
  • 13
    • 0344277931 scopus 로고    scopus 로고
    • note
    • Three tandem copies of GFP were used in order to (i) enhance the fluorescence signal and (ii) slow down the import of the two proteins to facilitate detection of a difference in their nuclear import.
  • 14
    • 0345571955 scopus 로고    scopus 로고
    • note
    • 3 indicate there is a similar amount of each full-length protein and a minor population of degraded protein (8).
  • 15
    • 0345571954 scopus 로고    scopus 로고
    • note
    • SE4-GFP expressed under the control of the PHO4 promoter on a CEN/ARS plasmid was still cytoplasmic after 2 hours of phosphate starvation. In contrast, wild-type Pho4-GFP was imported into the nucleus in less than 30 min after phosphate starvation (8). Additionally, a pse1-1 strain, defective for import of Pho4, cannot induce expression of acid phosphatase in response to phosphate starvation.
  • 16
    • 0345140353 scopus 로고    scopus 로고
    • note
    • SA1-GFP mutant has no defect in localization or regulation of acid phosphatase expression (8).
  • 17
    • 0345140354 scopus 로고    scopus 로고
    • note
    • SA4-GFP, which can be exported but whose import cannot be inhibited by phosphorylation, regulated acid phosphatase expression appropriately (8).
  • 18
    • 0026355413 scopus 로고    scopus 로고
    • note
    • SA1234PA6 does not undergo a shift in its electrophoretic mobility after incubation with Pho80/ Pho85 and adenosine 5′-triphosphate (ATP), suggesting that it cannot be phosphorylated (8).
  • 19
    • 0344709940 scopus 로고    scopus 로고
    • note
    • SA12346 is a weak activator of transcription (6).
  • 20
    • 0344709941 scopus 로고    scopus 로고
    • note
    • SA1234 also produces fully induced levels of acid phosphatase when grown in phosphate-rich medium (Fig. 3B). Thus, the additional mode of regulation requires Pho80, as expected if the regulation involves phosphorylation.
  • 21
    • 0028351946 scopus 로고    scopus 로고
    • note
    • PA6-GFP occurs with the same kinetics as wild-type Pho4 (8), suggesting that the ability of this mutant to interact with and be phosphorylated by Pho80 is similar to that of wild-type Pho4. (ii) Phosphorylation site 6 is in a domain involved in interaction with Pho2 (21), not with Pho80 (P. S. Jayaraman et al., above).
  • 25
    • 0344709937 scopus 로고    scopus 로고
    • note
    • SA1234PA6 interacted efficiently with Pho2 (Fig. 3C) (8) and activated transcription of acid phosphatase (Fig. 3B). Thus, Ala substitutions at Ser 1, 2, 3, and 4 do not affect the interaction with Pho2, and the Ser at site 6 is important for the interaction between Pho4 and Pho2.
  • 26
    • 0025072235 scopus 로고
    • All yeast strains are derived from K699 MATa [K. Nasmyth, G. Adolf, D. Lydall, A. Seddon, Cell 62, 631 (1990)]. PHO3 encodes an acid phosphatase expressed in phosphate-rich conditions [A. Toh-e, Y. Ueda, S.-I. Kakimoto, Y. Oshima, J. Bacteriol. 113, 727 (1973)].
    • (1990) Cell , vol.62 , pp. 631
    • Nasmyth, K.1    Adolf, G.2    Lydall, D.3    Seddon, A.4
  • 27
    • 0015578109 scopus 로고
    • All yeast strains are derived from K699 MATa [K. Nasmyth, G. Adolf, D. Lydall, A. Seddon, Cell 62, 631 (1990)]. PHO3 encodes an acid phosphatase expressed in phosphate-rich conditions [A. Toh-e, Y. Ueda, S.-I. Kakimoto, Y. Oshima, J. Bacteriol. 113, 727 (1973)].
    • (1973) J. Bacteriol. , vol.113 , pp. 727
    • Toh-E, A.1    Ueda, Y.2    Kakimoto, S.-I.3    Oshima, Y.4
  • 28
    • 0344709936 scopus 로고    scopus 로고
    • note
    • 4 was added to a final concentration of 20 mM. Photos were taken 5 to 10 min after the addition of phosphate.
  • 29
    • 0344709935 scopus 로고    scopus 로고
    • note
    • 6 (11).
  • 30
    • 0344709931 scopus 로고    scopus 로고
    • note
    • 3 (12) under the control of the GAL1-10 promoter was grown in synthetic raffinose medium at 30 °C to log phase. No fluorescence is visible under noninducing conditions when cells are grown in raffinose. Expression of each fusion protein was induced by addition of galactose to a final concentration of 2%, and localization was monitored as a function of time by fluorescence microscopy (11). The photo was taken 1.5 hours after the addition of galactose.
  • 31
    • 0021471049 scopus 로고    scopus 로고
    • note
    • 600 values.
  • 32
    • 0027169860 scopus 로고    scopus 로고
    • note
    • 2 elutions were run on 7.8% SDS-polyacrylamide gels and transferred to nitrocellulose membranes. Pho2 was detected by protein immunoblotting with polyclonal anti-Pho2.
  • 34
    • 0025354922 scopus 로고
    • P. C. McAndrew, J. Svaren, S. R. Martin, W. Hörz, C. R. Goding, Mol. Cell. Biol. 18, 5818 (1998); N. Ogawa and Y. Oshima, ibid. 10, 2224 (1990).
    • (1990) Mol. Cell. Biol. , vol.10 , pp. 2224
    • Ogawa, N.1    Oshima, Y.2
  • 35
    • 0345140350 scopus 로고    scopus 로고
    • note
    • 6-Gsp1Q71L; A. Kaffman for helpful discussions; and C. Gross, J. Weissman, R. Tjian, and members of the O'Shea lab for helpful comments on the manuscript Supported by the David and Lucile Packard Foundation (E.K.O.). A.K. was supported by a Howard Hughes Medical Institute Predoctoral Fellowship.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.