Live cell assays to identify regulators of ER to Golgi trafficking

LISAUSKAS, Tautvydas, Petr MATULA, Christoph CLAAS, Susanne REUSING, Stefan WIEMANN, Holger ERFLE, Lars LEHMANN, Peter FISCHER, Roland EILS, Karl ROHR, Brian STORRIE and Vytaute STARKUVIENE. Live cell assays to identify regulators of ER to Golgi trafficking. Traffic. John Wiley & Sons, 2012, vol. 13, No 3, p. 416–432. ISSN 1398-9219. Available from: https://dx.doi.org/10.1111/j.1600-0854.2011.01318.x.

Basic information

• Original name: Live cell assays to identify regulators of ER to Golgi trafficking
• Authors: LISAUSKAS, Tautvydas (440 Lithuania), Petr MATULA (203 Czech Republic, guarantor, belonging to the institution), Christoph CLAAS (276 Germany), Susanne REUSING (276 Germany), Stefan WIEMANN (276 Germany), Holger ERFLE (276 Germany), Lars LEHMANN (276 Germany), Peter FISCHER (276 Germany), Roland EILS (276 Germany), Karl ROHR (276 Germany), Brian STORRIE (840 United States of America) and Vytaute STARKUVIENE (440 Lithuania).
• Edition: Traffic, John Wiley & Sons, 2012, 1398-9219.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 4.652
• RIV identification code: RIV/00216224:14330/12:00059140
• Organization unit: Faculty of Informatics
• Doi: http://dx.doi.org/10.1111/j.1600-0854.2011.01318.x
• UT WoS: 000300054100006
• Keywords in English: BFA; GalT; ER to Golgi trafficking; YIPF; GOT1B; USE1; SACM1L
• Tags: International impact, Reviewed

Abstract

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.

Links

• MSM0021622419, plan (intention): Name: Vysoce paralelní a distribuované výpočetní systémy

Investor: Ministry of Education, Youth and Sports of the CR, Highly Parallel and Distributed Computing Systems

• 2B06052, research and development project: Name: Vytipování markerů, screening a časná diagnostika nádorových onemocnění pomocí vysoce automatizovaného zpracování multidimenzionálních biomedicínských obrazů (Acronym: Biomarker)

Investor: Ministry of Education, Youth and Sports of the CR, Determination of markers, screening and early diagnostics of cancer diseases using highly automated processing of multidimensional biomedical images