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This study underscores the possible role of phosphorylation at Ser\u2010256 in regulating AQP2 maturation. AQP2\u2010transfected renal CD8 cells were incubated with brefeldin A (BFA) to accumulate newly synthesized AQP2 in the endoplasmic reticulum (ER), and AQP2 flow from ER to the vesicular compartment was analyzed after BFA washout. We found that a) in the ER, AQP2 is weakly phosphorylated; b) the amount of phosphorylated AQP2 (p\u2010AQP2) at Ser\u2010256 increased significantly during transit in the Golgi, even in the presence of the PKA inhibitor H89; and c) AQP2 transport from the Golgi to the vasopressin\u2010regulated vesicular compartment occurred with a concomitant decrease in p\u2010AQP2 at Ser\u2010256. These results support the hypothesis that AQP2 transition in the Golgi apparatus is associated with a PKA\u2010independent increase in AQP2 phosphorylation at Ser\u2010256. Conversely, impaired constitutive phosphorylation in a Golgiassociated compartment occurring in cells expressing mutated S256A\u2010AQP2 or E258K\u2010AQP2 causes phosphorylation\u2010defective AQP2 routing to lysosomes. This result might explain the molecular basis of the dominant form of nephrogenic diabetes insipidus caused by the mutation E258K\u2010AQP2, in which the phenotype is caused by an impaired routing of AQP2.<\/jats:p>","DOI":"10.1096\/fj.02-0870fje","type":"journal-article","created":{"date-parts":[[2003,8,4]],"date-time":"2003-08-04T20:40:05Z","timestamp":1060029605000},"page":"1-24","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":79,"title":["Ser\u2010256 phosphorylation dynamics of aquaporin 2 during maturation from the endoplasmic reticulum to the vesicular compartment in renal cells"],"prefix":"10.1096","volume":"17","author":[{"given":"Giuseppe","family":"Procino","sequence":"first","affiliation":[{"name":"Dipartimento di Fisiologia Generale ed Ambientale University of Bari  70126 Bari Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monica","family":"Carmosino","sequence":"additional","affiliation":[{"name":"Dipartimento di Fisiologia Generale ed Ambientale University of Bari  70126 Bari Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oriano","family":"Marin","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica Biologica University of Padova  35121 Padova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna M.","family":"Brunanti","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica Biologica University of Padova  35121 Padova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonella","family":"Contri","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica Biologica University of Padova  35121 Padova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lorenzo A.","family":"Pinna","sequence":"additional","affiliation":[{"name":"Dipartimento di Chimica Biologica University of Padova  35121 Padova Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberta","family":"Mannucci","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Clinica e Sperimentale Universita' di Perugia  06100 Perugia Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Soren","family":"Nielsen","sequence":"additional","affiliation":[{"name":"Department of Cell Biology Institute of Anatomy University of Aarhus  8000 Aarhus Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tae-Hwan","family":"Kwon","sequence":"additional","affiliation":[{"name":"Department of Physiology School of Medicine Dongguk University  Kyungju 780-714 Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria","family":"Svelto","sequence":"additional","affiliation":[{"name":"Dipartimento di Fisiologia Generale ed Ambientale University of Bari  70126 Bari Italy"},{"name":"Centro di Eccellenza di Genomica Comparata University of Bari  Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanna","family":"Valenti","sequence":"additional","affiliation":[{"name":"Dipartimento di Fisiologia Generale ed Ambientale University of Bari  70126 Bari Italy"},{"name":"Centro di Eccellenza di Genomica Comparata University of Bari  Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2003,8]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.1997.272.6.F816"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.23.14800"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.1999.276.2.F254"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajprenal.2000.278.1.F29"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.151.4.919"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.28.16952"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.17.10349"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.142.6.1399"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.26.14461"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.275.25.19050"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.10.5552"},{"key":"e_1_2_6_13_1","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1242\/jcs.113.11.1985","article-title":"The phosphatase inhibitor okadaic acid induces AQP2 translocation independently from AQP2 phosphorylation in renal collecting duct cells","volume":"113","author":"Valenti G.","year":"2000","journal-title":"J. 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