title: Application of Gaussia luciferase in bicistronic and non-conventional secretion reporter constructs.
abstract: BACKGROUND
  Secreted luciferases are highly useful bioluminescent reporters for cell-based assays and drug discovery. A variety of secreted luciferases from marine organisms have been described that harbor an N-terminal signal peptide for release along the classical secretory pathway. Here, we have characterized the secretion of Gaussia luciferase in more detail.
  
  
  RESULTS
  We describe three basic mechanisms by which GLUC can be released from cells: first, classical secretion by virtue of the N-terminal signal peptide; second, internal signal peptide-mediated secretion and third, non-conventional secretion in the absence of an N-terminal signal peptide. Non-conventional release of dNGLUC is not stress-induced, does not require autophagy and can be enhanced by growth factor stimulation. Furthermore, we have identified the golgi-associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1) as a suppressor of release of dNGLUC.
  
  
  CONCLUSIONS
  Due to its secretion via multiple secretion pathways GLUC can find multiple applications as a research tool to study classical and non-conventional secretion. As GLUC can also be released from a reporter construct by internal signal peptide-mediated secretion it can be incorporated in a novel bicistronic secretion system.
authors.forename: Christin
authors.initials: C
authors.lastname: Luft
authors.forename: Jamie
authors.initials: J
authors.lastname: Freeman
authors.forename: David
authors.initials: D
authors.lastname: Elliott
authors.forename: Nadia
authors.initials: N
authors.lastname: Al-Tamimi
authors.forename: Janos
authors.initials: J
authors.lastname: Kriston-Vizi
authors.forename: Jacob
authors.initials: J
authors.lastname: Heintze
authors.forename: Ida
authors.initials: I
authors.lastname: Lindenschmidt
authors.forename: Brian
authors.initials: B
authors.lastname: Seed
authors.affiliation: Medical Research Council, Laboratory for Moleclar and Cell Biology, University College London, Gower Street, London WC1E 6BT, UK. r.ketteler@ucl.ac.uk.
authors.forename: Robin
authors.initials: R
authors.lastname: Ketteler
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001671:Biochemistry / Q000379:methods*; D012634:Bodily Secretions; D005796:Genes / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics
chemical_list: D018528:ATP-Binding Cassette Transporters; D033942:Adaptor Proteins, Vesicular Transport; D001426:Bacterial Proteins; C407306:GGA adaptor proteins; C421008:GluC protein, Corynebacterium glutamicum; D021382:Protein Sorting Signals; D049409:Luciferases, Firefly