Pax4 paired domain mediates direct protein transduction into mammalian cells.

Pax4, a paired-box transcription factor, is a key regulator of pancreatic islet cell growth and differentiation. Here, we report for the first time that the Pax4 protein can permeate into various cell types including pancreatic islets. The paired domain of Pax4 serves as a novel protein transduction domain (PTD). The Pax4 protein can transduce in a dose- and time-dependent manner. The cellular uptake of Pax4 PTD can be completely blocked by heparin, whereas cytochalasin D and amiloride were partially effective in blocking the Pax4 protein entry. Transduced intact Pax4 protein functions similarly to the endogenous Pax4. It inhibits the Pax6 mediated transactivation and protects Min6 cells against TNFalpha-induced apoptosis. These data suggest that Pax4 protein transduction could be a safe and valuable strategy for protecting islet cell growth in culture from apoptosis and promoting islet cell differentiation.

[1]  N. Kobayashi,et al.  BETA2/NeuroD protein transduction requires cell surface heparan sulfate proteoglycans. , 2006, Human gene therapy.

[2]  Hui-ling Wu,et al.  A novel type of PTD, common helix-loop-helix motif, could efficiently mediate protein transduction into mammalian cells. , 2006, Biochemical and biophysical research communications.

[3]  Lu Shun,et al.  Role of Pax4 in Pdx1-VP16-mediated liver-to-endocrine pancreas transdifferentiation , 2006, Laboratory Investigation.

[4]  S. Bonner-Weir,et al.  BETA2/NeuroD protein can be transduced into cells due to an arginine- and lysine-rich sequence. , 2005, Diabetes.

[5]  S. Dowdy,et al.  Corrigendum to “Cationic TAT peptide transduction domain enters cells by macropinocytosis” [J. Control. Release 102 (2005) 247–253] , 2005 .

[6]  S. Bonner-Weir,et al.  Mechanism of PDX-1 protein transduction. , 2005, Biochemical and biophysical research communications.

[7]  B. Lebleu,et al.  Cellular Uptake of Unconjugated TAT Peptide Involves Clathrin-dependent Endocytosis and Heparan Sulfate Receptors* , 2005, Journal of Biological Chemistry.

[8]  C. Wollheim,et al.  Association of childhood type 1 diabetes mellitus with a variant of PAX4: possible link to beta cell regenerative capacity , 2005, Diabetologia.

[9]  Richard E. Miller,et al.  High glucose inhibits apoptosis induced by serum deprivation in vascular smooth muscle cells via upregulation of Bcl-2 and Bcl-xl. , 2005, Diabetes.

[10]  Steven F Dowdy,et al.  Cationic TAT peptide transduction domain enters cells by macropinocytosis. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[11]  J. Seelig,et al.  The cationic cell-penetrating peptide CPP(TAT) derived from the HIV-1 protein TAT is rapidly transported into living fibroblasts: optical, biophysical, and metabolic evidence. , 2005, Biochemistry.

[12]  B. Sosa-Pineda.  The gene Pax4 is an essential regulator of pancreatic β-cell development , 2004 .

[13]  C. Wollheim,et al.  The diabetes-linked transcription factor PAX4 promotes β-cell proliferation and survival in rat and human islets , 2004, The Journal of cell biology.

[14]  A. Beaudet,et al.  Acute toxicity after high-dose systemic injection of helper-dependent adenoviral vectors into nonhuman primates. , 2004, Human gene therapy.

[15]  C. Ricordi,et al.  DELIVERY OF PROTEINS AND PEPTIDES INTO LIVE CELLS BY MEANS OF PROTEIN TRANSDUCTION DOMAINS: POTENTIAL APPLICATION TO ORGAN AND CELL TRANSPLANTATION , 2004, Transplantation.

[16]  I. Kockum,et al.  Interaction and association analysis of a type 1 diabetes susceptibility locus on chromosome 5q11-q13 and the 7q32 chromosomal region in Scandinavian families. , 2004, Diabetes.

[17]  Steven F Dowdy,et al.  Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis , 2004, Nature Medicine.

[18]  A. Prochiantz,et al.  Transduction peptides: from technology to physiology , 2004, Nature Cell Biology.

[19]  Kurt Ballmer-Hofer,et al.  Antennapedia and HIV Transactivator of Transcription (TAT) “Protein Transduction Domains” Promote Endocytosis of High Molecular Weight Cargo upon Binding to Cell Surface Glycosaminoglycans* , 2003, Journal of Biological Chemistry.

[20]  M. Johansson,et al.  Cell surface adherence and endocytosis of protein transduction domains. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[21]  S. Bonner-Weir,et al.  PDX-1 protein containing its own antennapedia-like protein transduction domain can transduce pancreatic duct and islet cells. , 2003, Diabetes.

[22]  N. Takasu,et al.  PAX4 mutation (R121W) as a prodiabetic variant in Okinawans. , 2003, Biochemical and biophysical research communications.

[23]  T. Sanke,et al.  A missense mutation of Pax4 gene (R121W) is associated with type 2 diabetes in Japanese. , 2001, Diabetes.

[24]  C. Ricordi,et al.  Proteins linked to a protein transduction domain efficiently transduce pancreatic islets. , 2001, Diabetes.

[25]  Y. Ueda.  Activin A increases Pax4 gene expression in pancreatic β cell lines , 2000 .

[26]  S. Schwarze,et al.  In vivo protein transduction: intracellular delivery of biologically active proteins, compounds and DNA. , 2000, Trends in pharmacological sciences.

[27]  S. Herzig,et al.  Tissue-specific transcriptional activity of a pancreatic islet cell-specific enhancer sequence/Pax6-binding site determined in normal adult tissues in vivo using transgenic mice. , 1999, Molecular endocrinology.

[28]  P. Gruss,et al.  Pax6 is required for differentiation of glucagon-producing α-cells in mouse pancreas , 1997, Nature.

[29]  P. Gruss,et al.  The Pax4 gene is essential for differentiation of insulin-producing β cells in the mammalian pancreas , 1997, Nature.

[30]  G. Elliott,et al.  Intercellular Trafficking and Protein Delivery by a Herpesvirus Structural Protein , 1997, Cell.

[31]  T. Pollard,et al.  Effects of cytochalasin, phalloidin, and pH on the elongation of actin filaments. , 1991, Biochemistry.

[32]  M. Bretscher,et al.  Distinct endocytotic pathways in epidermal growth factor-stimulated human carcinoma A431 cells [published erratum appears in J Cell Biol 1990 Mar;110(3):859] , 1989, The Journal of cell biology.

[33]  J. Epstein,et al.  Getting your Pax straight: Pax proteins in development and disease. , 2002, Trends in genetics : TIG.

[34]  A. Estreicher,et al.  The pancreatic beta-cell-specific transcription factor Pax-4 inhibits glucagon gene expression through Pax-6 , 2002, Diabetologia.