ITSN1 and Ruk/CIN85 colocalized to clathrin-coated pits in MCF-7 cells

• Datum: 2009
• Hauptverfasser: Nikolaienko, O.V., Skrypkina, I.Ya., Tsyba, L.O., Drobot, L.B., Rynditch, A.V.
• Format: Artikel
• Sprache: English
• Veröffentlicht: Інститут молекулярної біології і генетики НАН України 2009
• Schriftenreihe: Вiopolymers and Cell
• Schlagworte: Короткі повідомлення
• Online Zugang: https://nasplib.isofts.kiev.ua/handle/123456789/152964
• Назва журналу: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Zitieren: ITSN1 and Ruk/CIN85 colocalized to clathrin-coated pits in MCF-7 cells / O.V. Nikolaienko, I.Ya. Skrypkina, L.O. Tsyba, L.B. Drobot, A.V. Rynditch // Вiopolymers and Cell. — 2009. — Т. 25, № 5. — С. 424-427. — Бібліогр.: 16 назв. — англ.

Activation of receptor tyrosine kinases (RTK) by corresponding ligands results not only in signal propagation, but also initiates a number of processes, such as clathrin-mediated endocytosis, which precisely regulate biological outcome. These processes are tightly controlled by coordinated action of a plethora of proteins – enzymes, scaffolds and inhibitory molecules. An example of an endocytic accessory protein that also functions in cell signaling is provided by intersectin 1 (ITSN1). Previously we have shown that ITSN1 forms a complex with adaptor protein Ruk/CIN85 and ubiquitin ligase Cbl-b, which are implicated in down regulation of RTK. The present study aimed to determine the subcellular localization of ITSN1-Ruk/CIN85 complexes relatively to clathrin light chain and Cbl-b. Methods. Transient transfection of MCF-7 breast adenocarcinoma cells with the constructs containing Omni-tagged intersectin 1 and clathrin light chain fused with mCherry fluorescent protein was utilized to determine subcellular localization by direct or indirect immunofluorescence. Results. We found that Ruk/CIN85-ITSN1 complexes partially colocalized with Cbl-b and clathrin light chain in MCF-7 cells. Conclusions. In our report we provide experimental evidence that ITSN1-Ruk/CIN85 complexes exist in pre-assembled state with Cbl-b and are targeted to clathrin-coated pits in MCF-7 cells.