Mixed lineage kinase 2 interacts with clathrin and influences clathrin-coated vesicle trafficking

Akbarzadeh, S; Ji, H; Frecklington, D; Marmy-Conus, N; Mok, YF; Bowes, L; Devereux, L; Linsenmeyer, M; Simpson, RJ; Dorow, DS

Author(s): Akbarzadeh, S; Ji, H; Frecklington, D; Marmy-Conus, N; Mok, YF; Bowes, L; Devereux, L; Linsenmeyer, M; Simpson, RJ; Dorow, DS;
Details: Publication Year 2002-09-27,Volume 277,Issue #39,Page 36280-36287
Journal Title: JOURNAL OF BIOLOGICAL CHEMISTRY
Publication Type: Journal Article
Abstract: Mixed lineage kinase 2 (MLK2) is a protein kinase that signals in the stress-activated Jun N-terminal kinase signal transduction pathway. We used immunoprecipitation and mass spectrometric analysis to identify MLK2-binding proteins in cell lines with inducible expression of green fluorescent protein-tagged MLK2. Here we report the identification of clathrin as a binding partner for MLK2 in both cultured cells and mammalian brain. We demonstrate that clathrin binding requires a motif (LLDMD) located near the MLK2 C terminus, which is similar to "clathrin box" motifs important for binding of clathrin coat assembly and accessory proteins to the clathrin heavy chain. A C-terminal fragment of MLK2 containing this motif binds strongly to clathrin, and mutation of the LLDMD sequence to LAAAD completely abrogates clathrin binding. We isolated clathrin-coated vesicles from green fluorescent protein-MLK2-expressing cells and from mouse brain lysates and found that MLK2 is enriched along with clathrin in these vesicles. In addition, we demonstrated that endogenous MLK2 co-immunoprecipitates with clathrin heavy chain from the vesicle-enriched fraction of mouse brain lysate. Furthermore, overexpression of MLK2 in cultured cells inhibits accumulation of labeled transferrin in recycling endosomes during receptor-mediated endocytosis. These findings suggest a role for MLK2 and the stress-signaling pathway at sites of clathrin activity in vesicle formation or trafficking.
Publisher: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Keywords: ACTIN-BINDING PROTEIN; ZIPPER-BEARING KINASE; LEUCINE-ZIPPER; MEDIATED ENDOCYTOSIS; TERMINAL DOMAIN; RECEPTOR ACTIVATION; SIGNALING PROTEINS; MOLECULAR-CLONING; SH3 DOMAINS; C-JUN
Publisher's Version: https://doi.org/10.1074/jbc.M204626200
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2002-09-27 12:00:00