Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11
Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie
Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie
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Research Article Cell biology Oncology

Epithelial-to-mesenchymal transition drives a pro-metastatic Golgi compaction process through scaffolding protein PAQR11

Abstract

Tumor cells gain metastatic capacity through a Golgi phosphoprotein 3–dependent (GOLPH3-dependent) Golgi membrane dispersal process that drives the budding and transport of secretory vesicles. Whether Golgi dispersal underlies the pro-metastatic vesicular trafficking that is associated with epithelial-to-mesenchymal transition (EMT) remains unclear. Here, we have shown that, rather than causing Golgi dispersal, EMT led to the formation of compact Golgi organelles with improved ribbon linking and cisternal stacking. Ectopic expression of the EMT-activating transcription factor ZEB1 stimulated Golgi compaction and relieved microRNA-mediated repression of the Golgi scaffolding protein PAQR11. Depletion of PAQR11 dispersed Golgi organelles and impaired anterograde vesicle transport to the plasma membrane as well as retrograde vesicle tethering to the Golgi. The N-terminal scaffolding domain of PAQR11 was associated with key regulators of Golgi compaction and vesicle transport in pull-down assays and was required to reconstitute Golgi compaction in PAQR11-deficient tumor cells. Finally, high PAQR11 levels were correlated with EMT and shorter survival in human cancers, and PAQR11 was found to be essential for tumor cell migration and metastasis in EMT-driven lung adenocarcinoma models. We conclude that EMT initiates a PAQR11-mediated Golgi compaction process that drives metastasis.

Authors

Xiaochao Tan, Priyam Banerjee, Hou-Fu Guo, Stephen Ireland, Daniela Pankova, Young-ho Ahn, Irodotos Michail Nikolaidis, Xin Liu, Yanbin Zhao, Yongming Xue, Alan R. Burns, Jonathon Roybal, Don L. Gibbons, Tomasz Zal, Chad J. Creighton, Daniel Ungar, Yanzhuang Wang, Jonathan M. Kurie

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Figure 2

Ectopic ZEB1 expression increases Golgi compaction.

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Ectopic ZEB1 expression increases Golgi compaction. (A) Confocal microgr...
(A) Confocal micrographs of Golgi (GM130, green) and nuclei (DAPI, magenta) in 393P cells stably transfected with empty (Vec) or ZEB1 expression vectors. Scale bars: 3 μm. Scatter plots show the normalized Golgi areas in 393P cells (left), 307P cells (middle), and HCC827 cells (right) stably transfected with ZEB1 or empty (Vec) expression vectors. Each dot represents values from a single cell. (B) Scatter plots show the average Golgi element numbers (left) and areas (right) in 393P cells stably transfected with ZEB1 or empty (Vec) expression vectors. Each dot represents values from a single cell. (C) Pseudocolored images of FRAP assays using the Golgi enzyme GalNAcT in 393P_vector cells (left) and 393P_ZEB1 cells (right) at time points indicated to the left of each row. The bleached regions of interest are indicated by arrowheads, and intensity levels are indicated by LUT bar (right). Scale bars: 3 μm. (D) The scatter plot shows the intensity recovery profile (%) after photobleaching. n = 20 cells per group. (E) Electron micrographs of representative cisternal stacks (arrows) in 393P_vector cells (left) and 393P_ZEB1 cells (right). Scale bar: 1 μm. Bar graphs show the mean numbers of cisternae per stack (left) and cisternal lengths (right). n > 10 cells per group. P values were determined using 2-tailed Student’s t test. Results were replicated (n ≥ 2 experiments).