Interaction of the Unc-51-like kinase and microtubule-associated protein light chain 3 related proteins in the brain: possible role of vesicular transport in axonal elongation.
We identified two mammalian ULK1 (Unc-51-like kinase involved in neurite extension) binding proteins by yeast two-hybrid screening. Both proteins showed high structural similarity to microtubule-associated protein (MAP) light chain 3 (LC3). One is identical to the Golgi-associated ATPase Enhancer of 16 kDa (GATE-16), an essential factor for intra-Golgi transport [39]. The other is identical to the gamma 2-subunit of GABA-A receptor associated protein (GABARAP) which has a possible role in receptor transport [46]. Using the yeast two-hybrid system and the in vitro GST pull-down assay, we found that the N-terminal proline/serine rich (PS) domain of ULK1 (amino acid 287-416) is required for ULK1-GATE-16 and ULK1-GABARAP protein interactions. However, the kinase activity of ULK1 affected neither ULK1-GATE-16 nor ULK1-GABARAP interaction. Immunohistochemical analysis using ULK1 and GABARAP antibodies showed that the ULK1 and the GABARAP proteins co-localized to many kind of neurons such as pyramidal cells of the hippocampus, mitral cells of the olfactory bulb, and Purkinje cells of the cerebellum. In HeLa cells, endogenous ULK1 and tagged GABARAP showed punctate structures in the cytosol, and were colocalized. These results suggest that the interaction of ULK1 and GABARAP is important to vesicle transport and axonal elongation in mammalian neurons.
Okazaki
N
N
Helix Research Institute, 1532-3 Yana, Kisarazu-city, Chiba 292-0812, Japan. okazaki@hri.co.jp
Yan
J
J
Yuasa
S
S
Ueno
T
T
Kominami
E
E
Masuho
Y
Y
Koga
H
H
Muramatsu
M
M
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; D001343:Autophagy / Q000502:physiology; D000071190:Autophagy-Related Protein 8 Family; D001369:Axons / Q000201:enzymology*; D001921:Brain / Q000166:cytology / Q000201:enzymology*; D019556:COS Cells; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins / Q000378:metabolism; D018871:Endoplasmic Reticulum, Smooth / Q000201:enzymology; D015870:Gene Expression / Q000502:physiology; D006056:Golgi Apparatus / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D020798:Two-Hybrid System Techniques
D048868:Adaptor Proteins, Signal Transducing; D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; D000071190:Autophagy-Related Protein 8 Family; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; C116741:GABARAP protein, human; C498698:GABARAPL2 protein, human; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; C112443:UNC-51 protein, C elegans; D017346:Protein Serine-Threonine Kinases