{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T21:25:17Z","timestamp":1790717117733,"version":"4.1.0"},"reference-count":52,"publisher":"Portland Press Ltd.","issue":"2","license":[{"start":{"date-parts":[[2019,7,25]],"date-time":"2019-07-25T00:00:00Z","timestamp":1564012800000},"content-version":"vor","delay-in-days":862,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,4,30]]},"abstract":"<jats:p>In higher eukaryotes, cell proliferation is regulated by class I phosphatidylinositol 3-kinase (PI3K), which transduces stimuli received from neighboring receptors by local generation of PtdIns(3,4,5)P3 in cellular membranes. PI3K is a heterodimeric protein consisting of a regulatory and a catalytic subunit (p85 and p110 respectively). Heterologous expression of p110\u03b1 in Saccharomyces cerevisiae leads to toxicity by conversion of essential PtdIns(4,5)P2 into futile PtdIns(3,4,5)P3, providing a humanized yeast model for functional studies on this pathway. Here, we report expression and functional characterization in yeast of all regulatory and catalytic human PI3K isoforms, and exploitation of the most suitable setting to functionally assay panels of tumor- and germ line-associated PI3K mutations, with indications to the limits of the system. The activity of p110\u03b1 in yeast was not compromised by truncation of its N-terminal adaptor-binding domain (ABD) or inactivation of the Ras-binding domain (RBD). In contrast, a cluster of positively charged residues at the C2 domain was essential. Expression of a membrane-driven p65\u03b1 oncogenic-truncated version of p85\u03b1, but not the full-length protein, led to enhanced activity of \u03b1, \u03b2, and \u03b4 p110 isoforms. Mutations impairing the inhibitory regulation exerted by the p85\u03b1 iSH2 domain on the C2 domain of p110\u03b1 yielded the latter non-responsive to negative regulation, thus reproducing this oncogenic mechanism in yeast. However, p85\u03b1 germ line mutations associated with short stature, hyperextensibility of joints and\/or inguinal hernia, ocular depression, Rieger anomaly, and teething delay (SHORT) syndrome did not increase PI3K activity in this model, supporting the idea that SHORT syndrome-associated p85\u03b1 mutations operate through mechanisms different from the canonical disruption of inhibitory p85\u2013p110 interactions typical of cancer.<\/jats:p>","DOI":"10.1042\/bsr20160258","type":"journal-article","created":{"date-parts":[[2017,2,1]],"date-time":"2017-02-01T01:10:44Z","timestamp":1485911444000},"update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Insights into the pathological mechanisms of p85\u03b1 mutations using a yeast-based phosphatidylinositol 3-kinase model"],"prefix":"10.1042","volume":"37","author":[{"given":"Mar\u00eda\u00a0D.","family":"Oliver","sequence":"first","affiliation":[{"name":"Dpto. de Microbiolog\u00eda II, Facultad de Farmacia, Universidad Complutense de Madrid and Instituto Ram\u00f3n y Cajal de Investigaciones Sanitarias (IRyCIS), 28040 Madrid, Spain"},{"name":"Centro de Investigaci\u00f3n Pr\u00edncipe Felipe, 46012 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa","family":"Fern\u00e1ndez-Acero","sequence":"additional","affiliation":[{"name":"Dpto. de Microbiolog\u00eda II, Facultad de Farmacia, Universidad Complutense de Madrid and Instituto Ram\u00f3n y Cajal de Investigaciones Sanitarias (IRyCIS), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Luna","sequence":"additional","affiliation":[{"name":"Biocruces Health Research Institute, 48903 Barakaldo, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel","family":"Rodr\u00edguez-Escudero","sequence":"additional","affiliation":[{"name":"Dpto. de Microbiolog\u00eda II, Facultad de Farmacia, Universidad Complutense de Madrid and Instituto Ram\u00f3n y Cajal de Investigaciones Sanitarias (IRyCIS), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda","family":"Molina","sequence":"additional","affiliation":[{"name":"Dpto. de Microbiolog\u00eda II, Facultad de Farmacia, Universidad Complutense de Madrid and Instituto Ram\u00f3n y Cajal de Investigaciones Sanitarias (IRyCIS), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Pulido","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n Pr\u00edncipe Felipe, 46012 Valencia, Spain"},{"name":"Biocruces Health Research Institute, 48903 Barakaldo, Spain"},{"name":"IKERBASQUE, Basque Foundation for Science, 48013 Bilbao, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V\u00edctor\u00a0J.","family":"Cid","sequence":"additional","affiliation":[{"name":"Dpto. de Microbiolog\u00eda II, Facultad de Farmacia, Universidad Complutense de Madrid and Instituto Ram\u00f3n y Cajal de Investigaciones Sanitarias (IRyCIS), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2017,3,15]]},"reference":[{"key":"2021111619095196400_B1","doi-asserted-by":"crossref","first-page":"234","DOI":"10.2174\/1568009611313030002","article-title":"Diverse mechanisms of AKT pathway activation in human malignancy","volume":"13","author":"Cheung","year":"2013","journal-title":"Curr. 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Signal."},{"key":"2021111619095196400_B38","doi-asserted-by":"crossref","first-page":"20258","DOI":"10.1073\/pnas.0902369106","article-title":"Regulation of Class IA PI 3-kinases: C2 domain-iSH2 domain contacts inhibit p85\/p110alpha and are disrupted in oncogenic p85 mutants","volume":"106","author":"Wu","year":"2009","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111619095196400_B39","article-title":"SHORT syndrome","volume-title":"In GeneReviews(R)","author":"Innes","year":"1993"},{"key":"2021111619095196400_B40","doi-asserted-by":"crossref","first-page":"re2","DOI":"10.1126\/scisignal.2002165","article-title":"Structural basis for activation and inhibition of class I phosphoinositide 3-kinases","volume":"4","author":"Vadas","year":"2011","journal-title":"Sci. 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U.S.A."},{"key":"2021111619095196400_B42","doi-asserted-by":"crossref","first-page":"4562","DOI":"10.1158\/0008-5472.CAN-04-4114","article-title":"Functional analysis of PIK3CA gene mutations in human colorectal cancer","volume":"65","author":"Ikenoue","year":"2005","journal-title":"Cancer Res."},{"key":"2021111619095196400_B43","doi-asserted-by":"crossref","first-page":"15547","DOI":"10.1073\/pnas.1009652107","article-title":"Cancer-derived mutations in the regulatory subunit p85alpha of phosphoinositide 3-kinase function through the catalytic subunit p110alpha","volume":"107","author":"Sun","year":"2010","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111619095196400_B44","doi-asserted-by":"crossref","first-page":"16996","DOI":"10.1073\/pnas.0908444106","article-title":"A frequent kinase domain mutation that changes the interaction between PI3Kalpha and the membrane","volume":"106","author":"Mandelker","year":"2009","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111619095196400_B45","doi-asserted-by":"crossref","first-page":"3655","DOI":"10.1038\/onc.2011.532","article-title":"Regulation of lipid binding underlies the activation mechanism of class IA PI3-kinases","volume":"31","author":"Hon","year":"2012","journal-title":"Oncogene"},{"key":"2021111619095196400_B46","doi-asserted-by":"crossref","first-page":"2652","DOI":"10.1073\/pnas.0712169105","article-title":"Helical domain and kinase domain mutations in p110alpha of phosphatidylinositol 3-kinase induce gain of function by different mechanisms","volume":"105","author":"Zhao","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111619095196400_B47","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1042\/BJ20070681","article-title":"Effects of oncogenic p110alpha subunit mutations on the lipid kinase activity of phosphoinositide 3-kinase","volume":"409","author":"Carson","year":"2008","journal-title":"Biochem. J."},{"key":"2021111619095196400_B48","doi-asserted-by":"crossref","first-page":"e1003895","DOI":"10.1371\/journal.pcbi.1003895","article-title":"Investigating the structure and dynamics of the PIK3CA wild-type and H1047R oncogenic mutant","volume":"10","author":"Gkeka","year":"2014","journal-title":"PLoS Comput. Biol."},{"key":"2021111619095196400_B49","doi-asserted-by":"crossref","first-page":"15259","DOI":"10.1073\/pnas.1205508109","article-title":"Oncogenic mutations mimic and enhance dynamic events in the natural activation of phosphoinositide 3-kinase p110alpha (PIK3CA)","volume":"109","author":"Burke","year":"2012","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021111619095196400_B50","doi-asserted-by":"crossref","first-page":"1401","DOI":"10.1172\/JCI84005","article-title":"PI3-kinase mutation linked to insulin and growth factor resistance in vivo","volume":"126","author":"Winnay","year":"2016","journal-title":"J. Clin. 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