Protein phosphatase-1 is a novel regulator of the interaction between IRBIT and the inositol 1,4,5-trisphosphate receptor.
IRBIT is an IP3R [IP3 (inositol 1,4,5-trisphosphate) receptor]-binding protein that competes with IP3 for binding to the IP3R. Phosphorylation of IRBIT is essential for the interaction with the IP3R. The unique N-terminal region of IRBIT, residues 1-104 for mouse IRBIT, contains a PEST (Pro-Glu-Ser-Thr) domain with many putative phosphorylation sites. In the present study, we have identified a well-conserved PP1 (protein phosphatase-1)-binding site preceeding this PEST domain which enabled the binding of PP1 to IRBIT both in vitro and in vivo. IRBIT emerged as a mediator of its own dephosphorylation by associated PP1 and, hence, as a novel substrate specifier for PP1. Moreover, IRBIT-associated PP1 specifically dephosphorylated Ser68 of IRBIT. Phosphorylation of Ser68 was required for subsequent phosphorylation of Ser71 and Ser74, but the latter two sites were not targeted by PP1. We found that phosphorylation of Ser71 and Ser74 were sufficient to enable inhibition of IP3 binding to the IP3R by IRBIT. Finally, we have shown that mutational inactivation of the docking site for PP1 on IRBIT increased the affinity of IRBIT for the IP3R. This pinpoints PP1 as a key player in the regulation of IP3R-controlled Ca2+ signals.
Devogelaere
Benoit
B
Laboratory of Molecular and Cellular Signalling, Department of Molecular Cell Biology, Katholieke Universiteit Leuven, Campus Gasthuisberg O/N1, B-3000 Leuven, Belgium.
Beullens
Monique
M
Sammels
Eva
E
Derua
Rita
R
Waelkens
Etienne
E
van Lint
Johan
J
Parys
Jan B
JB
Missiaen
Ludwig
L
Bollen
Mathieu
M
De Smedt
Humbert
H
D043383:Adenosylhomocysteinase / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D015220:Calcium Channels / Q000378:metabolism*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D010766:Phosphorylation; D011485:Protein Binding; D054645:Protein Phosphatase 1 / Q000502:physiology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; C025389:Itpr1 protein, mouse; D008562:Membrane Glycoproteins; D018160:Receptors, Cytoplasmic and Nuclear; D054645:Protein Phosphatase 1; D043383:Adenosylhomocysteinase; C510392:IRBIT protein, mouse