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 3

PAQR11 depletion impairs Golgi compaction.

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PAQR11 depletion impairs Golgi compaction. (A) The bar graph shows the r...
(A) The bar graph shows the results of qPCR analysis performed to validate Affymetrix profiles of Golgi-related genes in 393P_ZEB1 cells and 393P_vector cells (n = 3). Expression levels are relative to values in 393P_vector cells, which were set at 1.0. *P < 0.05; #P < 0.01; †P < 0.001. (B) Scatter plots show Golgi areas normalized to nuclear areas in 344SQ cells (left), 393P_ZEB1 cells (middle), and H1299 cells (right) stably transfected with 1 of 2 distinct PAQR11 shRNAs (shPAQR11#1 or shPAQR11#3) or scrambled control shRNA (shCTL). Each dot represents values from a single cell. (C) Pseudocolored images of the Golgi enzyme GalNAcT during FRAP assays on 344SQ cells stably expressing PAQR11 shRNA (shPAQR11, bottom panels) or scrambled shRNA (shCTL, top panels) at time points indicated under the images. The bleached regions of interest are indicated by arrowheads, and intensity levels are indicated by LUT bars (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 344SQ_shCTL cells (left) and 344SQ_shPAQR11 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 or ANOVA for comparisons between 2 groups or more than 2 groups, respectively. Results were replicated (n ≥ 2 experiments).