GFP-cDNA Live & G-Protein Coupled Receptors
Dynamics of fluorescent cytoplasmic proteins upon activation of NK2 receptor

These web pages present the videos recorded during a live-imaging screen in HEK293 cells. This was aimed at identifying novel cytoplasmic proteins implicated in signaling pathways of G-protein coupled receptors, based on a translocation criterion (1).

The plasmids transfected in cells during the screen were from the GFP-tagged human full-length ORF plasmid collection and encoded proteins that localize in the cytoplasm (2-3). This plasmid library is composed of two vectors for each cDNA, one with the cDNA encoding CFP in frame 5’ of the ORF, the other with the cDNA encoding YFP in frame 3’ of the ORF.

An extensive study of REDD1, one of the hit proteins can be found in (4).

  1. Lecat S, Matthes HW, de Mey J, Pepperkok R, Simpson JC and Galzi JL. (2015) « A fluorescent live imaging screening assay based on translocation criteria identifies novel cytoplasmic proteins implicated in GPCR signalling pathways». Molecular and Cellular Proteomics. May;14(5):1385-99 First Published on March 10, 2015, doi:10.1074/mcp.M114.046698 http://www.mcponline.org/content/early/2015/03/10/mcp.M114.046698.full.pdf.
  2. Simpson JC, Wellenreuther R, Poustka A, Pepperkok R and Wiemann S (2000) “Systematic subcellular localization of novel proteins identified by large-scale cDNA sequencing”. EMBO Rep. 1(3):287-92. EMBL GFP-cDNA localization project, http://gfp-cdna.embl.de.
  3. Stadler C, Rexhepaj E, Singan VR, Murphy RF, Pepperkok R, Uhlén M, Simpson JC, and Lundberg E. (2013) « Immunofluorescence and fluorescent-protein tagging show high correlation for protein localization in mammalian cells.” Nat Methods. 4:315-23. doi: 10.1038/nmeth.2377.
  4. Michel G, Matthes HW, Hachet-Haas M, El Baghdadi K, de Mey J, Pepperkok R, Simpson JC, Galzi JL and Lecat S. (2014) “Plasma membrane translocation of REDD1 governed by GPCRs contributes to mTORC1 activation” Journal of Cell Science, 127:773-787. doi:10.1242/jcs.136432.

To get access to the list of the screened proteins, press « Show all ».

To refine the Search, use keywords such as : Name, full-Name, cDNA ID, Swissprot ID, biological process.

Show all
e.g WDR91, WD repeat-containing protein 91, AL136940, Q9H062, transcription
34 result(s)
Id Name Full Name Biological Process Clones cDNA ID Swissprot ID Endogenous localisation in Stadler et al. 2013 Fusion Orientation Localisation in Simpson et al. 2000 Localisation during screening Comments during screening
183 CNOT8 CCR4-NOT transcription complex subunit 8 RNA-mediated gene silencing transcription and translation regulations htes3_15m15 AL122045 Q9UFF9 no IF CFP-ORF cyt/nuc cyt/weak nuc
184 CNOT8 CCR4-NOT transcription complex subunit 8 RNA-mediated gene silencing transcription and translation regulations htes3_15m15 AL122045 Q9UFF9 no IF ORF-YFP cyt/nuc cyt/weak nuc bad transfection
215 RPAP1 RNA polymerase II-associated protein 1 Transcription hmcf_1m11 AL117479 Q9UFS7 No IF CFP-ORF cyt cyt few agg
216 RPAP1 RNA polymerase II-associated protein 1 Transcription hmcf_1m11 AL117479 Q9UFS7 No IF ORF-YFP ND ND
217 CPSF3L Integrator complex subunit 11 Transcription htes3_23a19 AL136813 Q9H0F9 IF cyt but confirmed nuc/cyt ? CFP-ORF cyt/nuc/ves strong nuc low cyt
218 CPSF3L Integrator complex subunit 11 Transcription htes3_23a19 AL136813 Q5TA45 IF cyt but confirmed nuc/cyt ? ORF-YFP cyt/nuc/ves mainly nuc
219 CALCOCO1 calcium-binding and coil-coiled domain containing protein1 transcription co-activator wingless signaling pathway htes3_7p9 AL136895 Q9H090 IF cyt CFP-ORF cyt cyt 1 central dot
220 CALCOCO1 calcium-binding and coil-coiled domain containing protein1 transcription co-activator wingless signaling pathway htes3_7p9 AL136895 Q9H090 IF cyt ORF-YFP cyt cyt
221 NRBF2 Nuclear receptor-binding factor 2 transcription regulation hfbr2_64c16 AL136656 Q9H0S9 No IF CFP-ORF cyt cyt
222 NRBF2 Nuclear receptor-binding factor 2 transcription regulation hfbr2_64c16 AL136656 Q9H0S9 No IF ORF-YFP cyt ND
223 LBH Protein LBH transcription regulation hfkd2_1j9 AL136703 Q9H0Q1 IF ves/nucleoli not confirmed CFP-ORF cyt cyt
224 LBH Protein LBH transcription regulation hfkd2_1j9 AL136703 Q9H0Q1 IF ves/nucleoli not confirmed ORF-YFP cyt/nuc cyt weak nuc out of nucleoli
225 ELP3 Elongator complex protein 3 transcription regulation hamy2_8l9 AL834273 Q9BVF7 IF negative CFP-ORF cyt cyt + dots in some cells
226 ELP3 Elongator complex protein 3 transcription regulation hamy2_8l9 AL834273 Q9BVF7 IF negative ORF-YFP cyt cyt/ weak nuc
227 GPBP1 Vasculin transcription regulation htes3_30g17 AL136844 Q9H0D4 IF cyt/ves/nucleoli/PM CFP-ORF cyt/ves cyt/nuc + ves and ves in nuc
228 GPBP1 Vasculin transcription regulation htes3_30g17 AL136844 Q86WP2 IF cyt/ves/nucleoli/PM ORF-YFP cyt/ves cyt + 1 or 2 big dots (centrosome ?)
229 ZNF280D Zinc finger protein 280D transcription regulation hfbr2_2h10 AL136634 Q9H0U5 IF cyt/nuc not nucleoli/Golgi/centrosome CFP-ORF cyt cyt
230 ZNF280D Zinc finger protein 280D transcription regulation hfbr2_2h10 AL136634 Q9H0U5 ORF-YFP cyt cyt
231 PCBD2 Pterin-4-alpha-carbinolamine dehydratase 2 transcription regulation hfkd2_46k19 AL136721 Q9H0N5 IF PM confirmed cyt/PM ? CFP-ORF cyt/nuc cyt/nuc less in nuc
232 PCBD2 Pterin-4-alpha-carbinolamine dehydratase 2 transcription regulation hfkd2_46k19 AL136721 Q9H0N5 IF PM confirmed cyt/PM ? ORF-YFP cyt/nuc negative cyt + agg
233 IKBKG NEMO NF-kappa-B essential modulator transcription regulation akxq28i231435 BC012114 Q9Y6K9 IF cyt CFP-ORF dots in nuc cyt/nuc more in nuc dots in nuc
234 IKBKG NEMO NF-kappa-B essential modulator transcription regulation akxq28i231435 BC012114 Q9Y6K9 IF cyt ORF-YFP ves( / nuc) dots in nuc contamination mitochondria staining
235 ZNF34 Zinc finger protein 34 Transcription regulation hsni_12k23 AL833814 Q9BSZ0 no IF CFP-ORF cyt/nuc cyt/nuc
236 ZNF34 Zinc finger protein 34 Transcription regulation hsni_12k23 AL833814 Q9BSZ0 ORF-YFP cyt/nuc (more in nuc) spots in nuc cyt/nuc bad transfection
237 TSC22D3 TSC22 domain family protein3 transcription regulation ? hfkd2_3a9 AL110191 Q99576 CFP-ORF cyt cyt
238 TSC22D3 TSC22 domain family protein3 transcription regulation ? hfkd2_3a9 AL110191 Q99576 ORF-YFP cyt/nuc ND
239 UHRF1BP1L UHRF1-binding protein 1-like transcription regulation ? related to UHRF1 function ? htes3_26m15 AL136821 Q9H0F1 error in Table and swissprot ID AOJNW5 CFP-ORF cyt cyt
240 UHRF1BP1L UHRF1-binding protein 1-like transcription regulation ? related to UHRF1 function ? htes3_26m15 AL136821 Q9H0F1 error in Table and swissprot ID AOJNW5 ORF-YFP cyt/nuc no transfected cells
241 FAM58A Cyclin-related protein FAM58A transcription regulation cell cycle ? akxq28d201286 BC032121 Q8N1B3 no IF CFP-ORF cyt/nuc + more in nuc +( speckles in nuc)/ves cyt/nuc more in nuc+spots
242 FAM58A Cyclin-related protein FAM58A transcription regulation cell cycle ? akxq28d201286 BC032121 Q8N1B3 No IF ORF-YFP cyt/nuc + more in nuc + speckles in nuc/ves speckles in nuc Not determined
243 BIRC5 Baculoviral IAP repeat-containing protein 5 transcription regulation cell cycle mitosis IMAGp958I1955 BC008718 O15392 IF cyt /nuc not nucleoli CFP-ORF cyt cyt
244 BIRC5 Baculoviral IAP repeat-containing protein 5 transcription regulation cell cycle mitosis IMAGp958I1955 BC008718 O15392 IF cyt /nuc not nucleoli ORF-YFP cyt cyt
245 TCF12 HTF4 Transcription factor 12 transcription regulation differentiation hlcc3_52k13 BX537967 Q7Z3D9 IF mitochondria not confirmed CFP-ORF cyt/nuc (less in nuc ) cyt/weak nuc + strong medium dots some perinuc
246 TCF12 HTF4 Transcription factor 12 transcription regulation differentiation hlcc3_52k13 BX537967 Q7Z3D9 IF mitochondria not confirmed ORF-YFP cyt/nuc strange :cyt/nuc + ER? or CK?
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