JLP associates with kinesin light chain 1 through a novel leucine zipper-like domain.
Scaffolding proteins exist in eukaryotes to properly assemble signaling proteins into specific multimeric functional complexes. JLP is a novel leucine zipper protein belonging to a family of scaffolding proteins that assemble JNK signaling modules. JLP is a proline-rich protein that contains two leucine zipper domains and a highly conserved C-terminal domain. We have identified kinesin light chain 1 (KLC1) as a binding partner for the second leucine zipper domain of JLP using yeast two-hybrid screening. The interaction domain of KLC1 was mapped to its tetratripeptide repeat, which contains a novel leucine zipper-like domain that is crucial for the interaction with JLP. Mutations of Leu-280, Leu-287, Val-294, and Leu-301 within this domain of KLC1 disrupted its ability to associate with JLP. Immunofluorescence studies showed that JLP and KLC1 co-localized in the cytoplasm and that the localization of JLP was dependent on its second leucine zipper. Ectopic expression of a dominant negative form of KLC1 resulted in the mislocalization of endogenous JLP. Moreover, the association between JLP and KLC1 occurred in vivo and was important in the formation of ternary complex with JNK1. These results identify a novel protein-protein interaction between KLC1 and JLP that involves leucine zipper-like domains and support the role of motor proteins in the spatial regulation of signaling modules.
Fels Institute for Cancer Research and Molecular Biology, School of Medicine, Temple University, Philadelphia, Pennsylvania 19140, USA.
Quang
Q
Nguyen
Clement M
CM
Lee
Anh
A
Le
E Premkumar
EP
Reddy
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005799:Genes, Dominant; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D047468:Immunoprecipitation; D016547:Kinesins; D007930:Leucine / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008954:Models, Biological; D009154:Mutation; D041681:NIH 3T3 Cells; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012190:Retroviridae / Q000235:genetics; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques; D014633:Valine / Q000737:chemistry
D048868:Adaptor Proteins, Signal Transducing; D018076:DNA, Complementary; C000715695:Kns2 protein, mouse; D008869:Microtubule-Associated Proteins; D010455:Peptides; C468928:Spag9 protein, mouse; D011392:Proline; D005982:Glutathione Transferase; D016547:Kinesins; D007930:Leucine; D014633:Valine