Publication details

Live cell assays to identify regulators of ER to Golgi trafficking

Authors

LISAUSKAS Tautvydas MATULA Petr CLAAS Christoph REUSING Susanne WIEMANN Stefan ERFLE Holger LEHMANN Lars FISCHER Peter EILS Roland ROHR Karl STORRIE Brian STARKUVIENE Vytaute

Year of publication 2012
Type Article in Periodical
Magazine / Source Traffic
MU Faculty or unit

Faculty of Informatics

Citation
Doi http://dx.doi.org/10.1111/j.1600-0854.2011.01318.x
Field Morphological specializations and cytology
Keywords BFA; GalT; ER to Golgi trafficking; YIPF; GOT1B; USE1; SACM1L
Description We used fluorescence microscopy based quantitative assays in living cells to identify regulators of ER to Golgi trafficking and/or Golgi complex maintenance. We first validated an automated procedure to identify factors, which influence Golgi to ER re-localization of GalT-CFP after BFA addition and wash-out. We then tested 14 proteins that potentially play a role in ER to Golgi trafficking, and localize to the ER and/or Golgi complex when over-expressed. 9 of them interfered with the rate of BFA induced redistribution of GalT-CFP to the ER, 6 of them interfered with GalT-CFP reassembly rate to a juxtanuclear region after BFA wash-out, and 6 of them were positive effectors in both assays. Notably, our live cell approach captures functions of those regulators of ER to Golgi trafficking, which were missed in previous fixed cell assays; as well as assigns respective roles for yet incompletely characterized proteins. Moreover, the assays can be extended to work under the conditions of RNAi and for testing chemical compounds.
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