3 Publications
 Next
3.1 Papers
Section Intro | Molecular modeling | Bioinformatics | docking | Methods and algorithms | Applications | Chronological list

3.1.6 Chronological list
Section Contents | ref 1 | ref 2 | ref 3 | ref 4 | ref 5 | ref 6 | ref 7 | ref 8 | ref 9 | ref 10 | ref 11 | ref 12 | ref 13 | ref 14 | ref 15 | ref 16 | ref 17 | ref 18 | ref 19 | ref 20 | ref 21 | ref 22 | ref 23 | ref 24 | ref 25 | ref 26 | ref 27 | ref 28 | ref 29 | ref 30 | ref 31 | ref 32 | ref 33 | ref 34 | ref 35 | ref 36 | ref 37 | ref 38 | ref 39 | ref 40 | ref 41 | ref 42 | ref 43 | ref 44 | ref 45 | ref 46 | ref 47 | ref 48 | ref 49 | ref 50 | ref 51 | ref 52 | ref 53 | ref 54 | ref 55 | ref 56 | ref 57 | ref 58 | ref 59 | ref 60 | ref 61 | ref 62 | ref 63 | ref 64 | ref 65 | ref 66 | ref 67 | ref 68 | ref 69 | ref 70 | ref 71 | ref 72 | ref 73 | ref 74 | ref 75 | ref 76 | ref 77 | ref 78 | ref 79 | ref 80 | ref 81 | ref 82 | ref 83 | in press

3.1.6.57 Isakoff, S.J., Cardozo, T., Andreev, J., Li, Z., Ferguson, K.M., Abagyan, R., Lemmon, M.A., Aronheim, A and Skolnik, E.Y. (1998). Identification and analysis of PH domain-containing targets of phosphatidylinositol 3-kinase using a novel in vivo assay in yeast. Embo J. 17 (18), 5374-87

Phosphatidylinositol 3-kinase (PI3K) mediates a variety of cellular responses by generating PtdIns(3,4)P2 and PtdIns(3,4,5)P3. These 3-phosphoinositides then function directly as second messengers to activate downstream signaling molecules by binding pleckstrin homology (PH) domains in these signaling molecules. We have established a novel assay in the yeast Saccharomyces cerevisiae to identify proteins that bind PtdIns(3,4)P2 and PtdIns(3,4,5)P3 in vivo which we have called TOPIS (Targets of PI3K Identification System). The assay uses a plasma membrane-targeted Ras to complement a temperature-sensitive CDC25 Ras exchange factor in yeast. Coexpression of PI3K and a fusion protein of activated Ras joined to a PH domain known to bind PtdIns(3,4)P2 (AKT) or PtdIns(3,4,5)P3 (BTK) rescues yeast growth at the non-permissive temperature of 37 degreesC. Using this assay, we have identified several amino acids in the beta1-beta2 region of PH domains that are critical for high affinity binding to PtdIns(3,4)P2 and/or PtdIns(3,4,5)P3, and we have proposed a structural model for how these PH domains might bind PI3K products with high affinity. From these data, we derived a consensus sequence which predicts high-affinity binding to PtdIns(3, 4)P2 and/or PtdIns(3,4,5)P3, and we have identified several new PH domain-containing proteins that bind PI3K products, including Gab1, Dos, myosinX, and Sbf1. Use of this assay to screen for novel cDNAs which rescue yeast at the non-permissive temperature should provide a powerful approach for uncovering additional targets of PI3K.