Details

Autor(en) / Beteiligte

• Wetzel, Franziska
• Mittag, Sonnhild
• Cano-Cortina, Misael
• Wagner, Tobias
• Krämer, Oliver H.
• Niedenthal, Rainer
• Gonzalez-Mariscal, Lorenza
• Huber, Otmar

Titel

SUMOylation regulates the intracellular fate of ZO-2

Ist Teil von

• Cellular and molecular life sciences : CMLS, 2017-01, Vol.74 (2), p.373-392

Ort / Verlag

Cham: Springer International Publishing

Erscheinungsjahr

2017

Link zum Volltext

Quelle

Springer journals

Beschreibungen/Notizen

• The zonula occludens (ZO)-2 protein links tight junctional transmembrane proteins to the actin cytoskeleton and associates with splicing and transcription factors in the nucleus. Multiple posttranslational modifications control the intracellular distribution of ZO-2. Here, we report that ZO-2 is a target of the SUMOylation machinery and provide evidence on how this modification may affect its cellular distribution and function. We show that ZO-2 associates with the E2 SUMO-conjugating enzyme Ubc9 and with SUMO-deconjugating proteases SENP1 and SENP3. In line with this, modification of ZO-2 by endogenous SUMO1 was detectable. Ubc9 fusion-directed SUMOylation confirmed SUMOylation of ZO-2 and was inhibited in the presence of SENP1 but not by an enzymatic-dead SENP1 protein. Moreover, lysine 730 in human ZO-2 was identified as a potential modification site. Mutation of this site to arginine resulted in prolonged nuclear localization of ZO-2 in nuclear recruitment assays. In contrast, a construct mimicking constitutive SUMOylation of ZO-2 (SUMO1ΔGG-ZO-2) was preferentially localized in the cytoplasm. Based on previous findings the differential localization of these ZO-2 constructs may affect glycogen-synthase-kinase-3β (GSK3β) activity and β-catenin/TCF-4-mediated transcription. In this context we observed that ZO-2 directly binds to GSK3β and SUMO1ΔGG-ZO-2 modulates its kinase activity. Moreover, we show that ZO-2 forms a complex with β-catenin. Wild-type ZO-2 and ZO-2-K730R inhibited transcriptional activity in reporter gene assays, whereas the cytosolic SUMO1ΔGG-ZO-2 did not. From these data we conclude that SUMOylation affects the intracellular localization of ZO-2 and its regulatory role on GSK3β and β-catenin signaling activity.