TY - JOUR
T1 - A specific dileucine motif is required for the GGA-dependent entry of newly synthesized insulin-responsive aminopeptidase into the insulin-responsive compartment
AU - Hou, June Chunqiu
AU - Suzuki, Naoko
AU - Pessin, Jeffrey E.
AU - Watson, Robert T.
PY - 2006/11/3
Y1 - 2006/11/3
N2 - In muscle and adipose cells, the insulin-responsive aminopeptidase (IRAP) is localized to intracellular storage sites and undergoes insulin-dependent redistribution to the cell surface. Following expression, the newly synthesized IRAP protein traffics to the perinuclear insulin-sensitive compartment and acquires insulin sensitivity 6-9 h following biosynthesis. Knockdown of GGA1 by RNA interference prevented IRAP from entering, but not exiting, the insulin-responsive compartment. Mutation of the dileucine motif at positions 76 and 77 (EGFP-IRAP/AA76,77), but not the dileucine motif at positions 53 and 54, resulted in the rapid default of the reporter to the cell surface beginning at 3 h following biosynthesis. Alanine substitution of 9 residues amino- or carboxyl-terminal to LL76,77 did not perturb basal intracellular sequestration or abrogate insulin-stimulated IRAP translocation. Moreover, a dominant interfering GGA mutant (VHS-GAT) potently inhibited insulin-stimulated translocation of EGFP-IRAP/WT but did not block the constitutive exocytotic trafficking of EGFP-IRAP/AA76,77. In addition, the EGFP-IRAP/WT and EGFP-IRAP/AA76,77 constructs occupied morphologically distinct tubulovesicular compartments in the perinuclear region. Taken together, these data indicate that LL76,77 functions during the GGA-dependent sorting of newly made IRAP into the insulin-responsive storage compartment.
AB - In muscle and adipose cells, the insulin-responsive aminopeptidase (IRAP) is localized to intracellular storage sites and undergoes insulin-dependent redistribution to the cell surface. Following expression, the newly synthesized IRAP protein traffics to the perinuclear insulin-sensitive compartment and acquires insulin sensitivity 6-9 h following biosynthesis. Knockdown of GGA1 by RNA interference prevented IRAP from entering, but not exiting, the insulin-responsive compartment. Mutation of the dileucine motif at positions 76 and 77 (EGFP-IRAP/AA76,77), but not the dileucine motif at positions 53 and 54, resulted in the rapid default of the reporter to the cell surface beginning at 3 h following biosynthesis. Alanine substitution of 9 residues amino- or carboxyl-terminal to LL76,77 did not perturb basal intracellular sequestration or abrogate insulin-stimulated IRAP translocation. Moreover, a dominant interfering GGA mutant (VHS-GAT) potently inhibited insulin-stimulated translocation of EGFP-IRAP/WT but did not block the constitutive exocytotic trafficking of EGFP-IRAP/AA76,77. In addition, the EGFP-IRAP/WT and EGFP-IRAP/AA76,77 constructs occupied morphologically distinct tubulovesicular compartments in the perinuclear region. Taken together, these data indicate that LL76,77 functions during the GGA-dependent sorting of newly made IRAP into the insulin-responsive storage compartment.
UR - http://www.scopus.com/inward/record.url?scp=33845465217&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33845465217&partnerID=8YFLogxK
U2 - 10.1074/jbc.M601583200
DO - 10.1074/jbc.M601583200
M3 - Article
C2 - 16945927
AN - SCOPUS:33845465217
SN - 0021-9258
VL - 281
SP - 33457
EP - 33466
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 44
ER -