The amino terminus extension in the long dipeptidyl peptidase 9 isoform contains a nuclear localization signal targeting the active peptidase to the nucleus

Cell Mol Life Sci. 2014 Sep;71(18):3611-26. doi: 10.1007/s00018-014-1591-6. Epub 2014 Feb 23.


The intracellular prolyl peptidase DPP9 is implied to be involved in various cellular pathways including amino acid recycling, antigen maturation, cellular homeostasis, and viability. Interestingly, the major RNA transcript of DPP9 contains two possible translation initiation sites, which could potentially generate a longer (892 aa) and a shorter version (863 aa) of DPP9. Although the endogenous expression of the shorter DPP9 form has been previously verified, it is unknown whether the longer version is expressed, and what is its biological significance. By developing specific antibodies against the amino-terminal extension of the putative DPP9-long form, we demonstrate for the first time the endogenous expression of this longer isoform within cells. Furthermore, we show that DPP9-long represents a significant fraction of total DPP9 in cells, under steady-state conditions. Using biochemical cell fractionation assays in combination with immunofluorescence studies, we find the two isoforms localize to separate subcellular compartments. Whereas DPP9-short is present in the cytosol, DPP9-long localizes preferentially to the nucleus. This differential localization is attributed to a classical monopartite nuclear localization signal (K(K/R)X(K/R)) in the N-terminal extension of DPP9-long. Furthermore, we detect prolyl peptidase activity in nuclear fractions, which can be inhibited by specific DPP8/9 inhibitors. In conclusion, a considerable fraction of DPP9, which was previously considered as a purely cytosolic peptidase, localizes to the nucleus and is active there, raising the intriguing possibility that the longer DPP9 isoform may regulate the activity or stability of nuclear proteins, such as transcription factors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Nucleus / metabolism*
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / analysis
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / chemistry*
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / metabolism
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / physiology
  • HeLa Cells
  • Humans
  • Isoenzymes / chemistry
  • Isoenzymes / metabolism
  • Isoenzymes / physiology
  • Molecular Sequence Data
  • Nuclear Localization Signals*
  • Protein Transport


  • Isoenzymes
  • Nuclear Localization Signals
  • DPP9 protein, human
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases