Application of Gaussia luciferase in bicistronic and non-conventional secretion reporter constructs

BMC Biochem. 2014 Jul 9:15:14. doi: 10.1186/1471-2091-15-14.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / metabolism*
  • Adaptor Proteins, Vesicular Transport / genetics
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biochemistry / methods*
  • Bodily Secretions
  • Genes / genetics
  • Genes, Reporter / genetics
  • HEK293 Cells
  • Humans
  • Luciferases, Firefly / genetics
  • Luciferases, Firefly / metabolism*
  • Protein Sorting Signals / genetics

Substances

  • ATP-Binding Cassette Transporters
  • Adaptor Proteins, Vesicular Transport
  • Bacterial Proteins
  • GGA adaptor proteins
  • GluC protein, Corynebacterium glutamicum
  • Protein Sorting Signals
  • Luciferases, Firefly