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Volumn 285, Issue 5430, 1999, Pages 1084-1087

Control of the terminal step of intracellular membrane fusion by protein phosphatase

Author keywords

[No Author keywords available]

Indexed keywords

PHOSPHOPROTEIN PHOSPHATASE 1;

EID: 0033551865     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5430.1084     Document Type: Article
Times cited : (139)

References (41)
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    • +) were generated by crossing this strain with PAY700-4 and PAY704-1, respectively, then selecting haploid segregants with the appropriate combination of genetic markers after tetrad analysis. PAY821-2Bα (MATα glc7::LEU2 trp1:: glc7-10::TRP1 pep4Δ::URA3) and PAY820-1Dα (MATα glc7::LEU2 trp1::GLC7::TRP1 pep4Δ::URA3) were similarly generated after crossing WZY333 (MATαpep4Δ:: URA3) with PAY700-4 and PAY704-1 and additionally verified by using a standard plate assay to confirm the lack of CPY activity after replating several times (31).
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    • Gdi1p extracts the Rab-like GTPase Ypt7p from the membrane and prevents docking (7). Anti-PP1 was not as effective in this type of experiment as in Fig. 1B because the antibody binds slowly to PP1 on the membrane (4). Because the kinetic analysis in Fig. 3 did not allow a preincubation of vacuoles with the antibodies as in Fig. 1B, the inhibitory effect of the antibodies was not complete, shifting the inhibition curve to the left. Microcystin LR binds covalently to PP1, resulting in rapid inactivation.
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    • We thank C. Macintosh for helpful hints in the initial phase of the project, K. Nasmyth for strain WZY333, M. Bayer and O. Müller for discussions, and E. Schlegel for help in preparing the manuscript. Supported by the Wellcome Trust (project grants 046956 and 049778, M.J.R.S.), the Deutsche Forschungsgemeinschaft (grant SFB445, A.M.), and the Boehringer Ingelheim Stiftung (A.M.).


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