BML-265 and Tyrphostin AG1478 Disperse the Golgi Apparatus and Abolish Protein Transport in Human Cells

  • Front Cell Dev Biol. 2019 Oct 11:7:232. doi: 10.3389/fcell.2019.00232.
Gaelle Boncompain  1 ,  Nelly Gareil  1 ,  Sarah Tessier  2 ,  Aurianne Lescure  2 ,  Thouis R Jones  2 ,  Oliver Kepp  3  4 ,  Guido Kroemer  3  4  5  6  7 ,  Elaine Del Nery  2 ,  Franck Perez  1
Affiliations
  • 1. Dynamics of Intracellular Organization Laboratory, Institut Curie, PSL Research University, Sorbonne Université, Centre National de la Recherche Scientifique, UMR 144, Paris, France.
  • 2. BioPhenics High-Content Screening Laboratory, Cell and Tissue Imaging Facility (PICT-IBiSA), Institut Curie, PSL Research University, Translational Research Department, Paris, France.
  • 3. Equipe Labellisée par la Ligue Contre le Cancer, Université de Paris, Sorbonne Université, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France.
  • 4. Metabolomics and Cell Biology Platforms, Gustave Roussy, Villejuif, France.
  • 5. Suzhou Institute for Systems Medicine, Chinese Academy of Medical Sciences, Suzhou, China.
  • 6. Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.
  • 7. Department of Women's and Children's Health, Karolinska University Hospital, Karolinska Institute, Stockholm, Sweden.
PMID: 31681765 DOI: 10.3389/fcell.2019.00232
Abstract

The steady-state localization of Golgi-resident glycosylation Enzymes in the Golgi apparatus depends on a balance between anterograde and retrograde transport. Using the Retention Using Selective Hooks (RUSH) assay and high-content screening, we identified small molecules that perturb the localization of Mannosidase II (ManII) used as a model cargo for Golgi resident Enzymes. In particular, we found that two compounds known as EGFR tyrosine kinase inhibitors, namely BML-265 and Tyrphostin AG1478 disrupt Golgi integrity and abolish secretory protein transport of diverse cargos, thus inducing brefeldin A-like effects. Interestingly, BML-265 and Tyrphostin AG1478 affect Golgi integrity and transport in human cells but not in rodent cells. The effects of BML-265 are reversible since Golgi integrity and protein transport are quickly restored upon washout of the compounds. BML-265 and Tyrphostin AG1478 do not lead to endosomal tubulation suggesting that, contrary to brefeldin A, they do not target the trans-Golgi ARF GEF BIG1 and BIG2. They quickly induce COPI dissociation from Golgi membranes suggesting that, in addition to EGFR kinase, the cis-Golgi ARF GEF GBF1 might also be a target of these molecules. Accordingly, overexpression of GBF1 prevents the effects of BML-265 and Tyrphostin AG1478 on Golgi integrity.

Keywords
EGFR kinase inhibitor; GBF1; golgi; high-content screening; membrane trafficking.
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-169948
    BML-265
    EGFR Tyrosine Kinase Inhibitor
    target: EGFR
    Research Areas: Cancer