Mammalian Golgi-associated Bicaudal-D2 functions in the dynein-dynactin pathway by interacting with these complexes.
Genetic analysis in Drosophila suggests that Bicaudal-D functions in an essential microtubule-based transport pathway, together with cytoplasmic dynein and dynactin. However, the molecular mechanism underlying interactions of these proteins has remained elusive. We show here that a mammalian homologue of Bicaudal-D, BICD2, binds to the dynamitin subunit of dynactin. This interaction is confirmed by mass spectrometry, immunoprecipitation studies and in vitro binding assays. In interphase cells, BICD2 mainly localizes to the Golgi complex and has properties of a peripheral coat protein, yet it also co-localizes with dynactin at microtubule plus ends. Overexpression studies using green fluorescent protein-tagged forms of BICD2 verify its intracellular distribution and co-localization with dynactin, and indicate that the C-terminus of BICD2 is responsible for Golgi targeting. Overexpression of the N-terminal domain of BICD2 disrupts minus-end-directed organelle distribution and this portion of BICD2 co-precipitates with cytoplasmic dynein. Nocodazole treatment of cells results in an extensive BICD2-dynactin-dynein co-localization. Taken together, these data suggest that mammalian BICD2 plays a role in the dynein- dynactin interaction on the surface of membranous organelles, by associating with these complexes.
Hoogenraad
C C
CC
MGC Department of Cell Biology, Erasmus University, PO Box 1738, 3000 DR Rotterdam, The Netherlands.
Akhmanova
A
A
Howell
S A
SA
Dortland
B R
BR
De Zeeuw
C I
CI
Willemsen
R
R
Visser
P
P
Grosveld
F
F
Galjart
N
N
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D004331:Drosophila melanogaster; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008322:Mammals; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D012441:Saccharomyces cerevisiae; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
C435206:Bicd2 protein, mouse; D002352:Carrier Proteins; C000606361:DCTN2 protein, human; D018076:DNA, Complementary; C000606365:Dctn2 protein, mouse; D000072159:Dynactin Complex; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D004398:Dyneins; D015739:Nocodazole