Depletion of kinesin 5B affects lysosomal distribution and stability and induces peri-nuclear accumulation of autophagosomes in cancer cells.
BACKGROUND
Enhanced lysosomal trafficking is associated with metastatic cancer. In an attempt to discover cancer relevant lysosomal motor proteins, we compared the lysosomal proteomes from parental MCF-7 breast cancer cells with those from highly invasive MCF-7 cells that express an active form of the ErbB2 (DeltaN-ErbB2).


RESULTS
Mass spectrometry analysis identified kinesin heavy chain protein KIF5B as the only microtubule motor associated with the lysosomes in MCF-7 cells, and ectopic DeltaN-ErbB2 enhanced its lysosomal association. KIF5B associated with lysosomes also in HeLa cervix carcinoma cells as analyzed by subcellular fractionation. The depletion of KIF5B triggered peripheral aggregations of lysosomes followed by lysosomal destabilization, and cell death in HeLa cells. Lysosomal exocytosis in response to plasma membrane damage as well as fluid phase endocytosis functioned, however, normally in these cells. Both HeLa and MCF-7 cells appeared to express similar levels of the KIF5B isoform but the death phenotype was weaker in KIF5B-depleted MCF-7 cells. Surprisingly, KIF5B depletion inhibited the rapamycin-induced accumulation of autophagosomes in MCF-7 cells. In KIF5B-depleted cells the autophagosomes formed and accumulated in the close proximity to the Golgi apparatus, whereas in the control cells they appeared uniformly distributed in the cytoplasm.


CONCLUSIONS
Our data identify KIF5B as a cancer relevant lysosomal motor protein with additional functions in autophagosome formation.
Cardoso
Carla M P
CM
Center for Neuroscience and Cell Biology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
Groth-Pedersen
Line
L
Høyer-Hansen
Maria
M
Kirkegaard
Thomas
T
Corcelle
Elizabeth
E
Andersen
Jens S
JS
Jäättelä
Marja
M
Nylandsted
Jesper
J
D001343:Autophagy; D016923:Cell Death; D045744:Cell Line, Tumor; D006801:Humans; D016547:Kinesins / Q000172:deficiency* / Q000502:physiology; D008247:Lysosomes / Q000378:metabolism*; D013058:Mass Spectrometry; D009369:Neoplasms / Q000473:pathology*; D015388:Organelles / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism*; D021381:Protein Transport; D040901:Proteomics; D018719:Erb-b2 Receptor Tyrosine Kinases; D020123:Sirolimus
D016547:Kinesins; D018719:Erb-b2 Receptor Tyrosine Kinases; D020123:Sirolimus; C488101:KIF5B protein, human; C508053:ERBB2 protein, human