SUMO-1 controls the protein stability and the biological function
of phosducin
C. Klenk, J. Humrich, U. Quitterer, and M. Lohse. J Biol Chem, 281 (13):
8357-64(March 2006)Klenk, Christoph Humrich, Jan Quitterer, Ursula Lohse, Martin J Comparative
Study Research Support, Non-U.S. Gov't United States The Journal
of biological chemistry J Biol Chem. 2006 Mar 31;281(13):8357-64.
Epub 2006 Jan 18..
Abstract
Phosducin regulates Gbetagamma-stimulated signaling by binding to
Gbetagamma subunits of heterotrimeric G-proteins. Control of phosducin
activity by phosphorylation is well established. However, little
is known about other mechanisms that may control phosducin activity.
Here we report that phosducin is regulated at the posttranslational
level by modification with the small ubiquitin-related modifier,
SUMO. We demonstrate modification with SUMO for phosducin in vitro
expressed in cells and for native phosducin purified from retina
and the heart. A consensus motif for SUMOylation was identified in
phosducin at amino acid positions 32-35. Mutation of the conserved
lysine 33 to arginine in this motif abolished SUMOylation of phosducin,
indicating that SUMO is attached to lysine 33 of phosducin. In transfected
cells the steady-state levels of the K33R mutant protein were much
lower compared with wild-type phosducin. The investigation of the
stability of wild-type phosducin and of phosducinK33R showed a decreased
protein stability of the SUMOylation-deficient mutant. The decreased
protein stability correlated with increased ubiquitinylation of the
SUMOylation-deficient mutant. These findings indicate that SUMOylation
protects phosducin from proteasomal degradation. SUMOylation of phosducin
decreased its ability to bind Gbetagamma. PhlP, a closely related
member of the phosducin family, was not a target for SUMOylation,
but its SUMOylation can be achieved by a single amino acid insertion
in the conserved N terminus of PhlP. Together, these findings show
that phosducin is a previously unrecognized target of SUMO modification
and that SUMOylation controls phosducin stability in cells as well
as its functional properties.
Klenk, Christoph Humrich, Jan Quitterer, Ursula Lohse, Martin J Comparative
Study Research Support, Non-U.S. Gov't United States The Journal
of biological chemistry J Biol Chem. 2006 Mar 31;281(13):8357-64.
Epub 2006 Jan 18.
%0 Journal Article
%1 Klenk2006
%A Klenk, C.
%A Humrich, J.
%A Quitterer, U.
%A Lohse, M. J.
%D 2006
%J J Biol Chem
%K & Acid Amino Animals Binding Blotting, COS Carrier Cell Cercopithecus Consensus Culture Data Eye G-Protein-Coupled/metabolism GTP-Binding Homology, Humans Line Lysine/chemistry Molecular Motifs Mutation Myocardium/chemistry Nerve Phosphoproteins/chemistry/genetics/isolation Phosphorylation Precipitin Protein Protein/genetics/*metabolism Proteins Proteins/chemistry/genetics/isolation Proteins/chemistry/metabolism Rats Recombinant Regulators Retina/chemistry Rhodopsin/metabolism SUMO-1 Sequence Sites Subunits/metabolism Techniques Tests Tissue Weight Western aethiops beta gamma purification/*metabolism Receptor
%N 13
%P 8357-64
%T SUMO-1 controls the protein stability and the biological function
of phosducin
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16421094
%V 281
%X Phosducin regulates Gbetagamma-stimulated signaling by binding to
Gbetagamma subunits of heterotrimeric G-proteins. Control of phosducin
activity by phosphorylation is well established. However, little
is known about other mechanisms that may control phosducin activity.
Here we report that phosducin is regulated at the posttranslational
level by modification with the small ubiquitin-related modifier,
SUMO. We demonstrate modification with SUMO for phosducin in vitro
expressed in cells and for native phosducin purified from retina
and the heart. A consensus motif for SUMOylation was identified in
phosducin at amino acid positions 32-35. Mutation of the conserved
lysine 33 to arginine in this motif abolished SUMOylation of phosducin,
indicating that SUMO is attached to lysine 33 of phosducin. In transfected
cells the steady-state levels of the K33R mutant protein were much
lower compared with wild-type phosducin. The investigation of the
stability of wild-type phosducin and of phosducinK33R showed a decreased
protein stability of the SUMOylation-deficient mutant. The decreased
protein stability correlated with increased ubiquitinylation of the
SUMOylation-deficient mutant. These findings indicate that SUMOylation
protects phosducin from proteasomal degradation. SUMOylation of phosducin
decreased its ability to bind Gbetagamma. PhlP, a closely related
member of the phosducin family, was not a target for SUMOylation,
but its SUMOylation can be achieved by a single amino acid insertion
in the conserved N terminus of PhlP. Together, these findings show
that phosducin is a previously unrecognized target of SUMO modification
and that SUMOylation controls phosducin stability in cells as well
as its functional properties.
@article{Klenk2006,
abstract = {Phosducin regulates Gbetagamma-stimulated signaling by binding to
Gbetagamma subunits of heterotrimeric G-proteins. Control of phosducin
activity by phosphorylation is well established. However, little
is known about other mechanisms that may control phosducin activity.
Here we report that phosducin is regulated at the posttranslational
level by modification with the small ubiquitin-related modifier,
SUMO. We demonstrate modification with SUMO for phosducin in vitro
expressed in cells and for native phosducin purified from retina
and the heart. A consensus motif for SUMOylation was identified in
phosducin at amino acid positions 32-35. Mutation of the conserved
lysine 33 to arginine in this motif abolished SUMOylation of phosducin,
indicating that SUMO is attached to lysine 33 of phosducin. In transfected
cells the steady-state levels of the K33R mutant protein were much
lower compared with wild-type phosducin. The investigation of the
stability of wild-type phosducin and of phosducinK33R showed a decreased
protein stability of the SUMOylation-deficient mutant. The decreased
protein stability correlated with increased ubiquitinylation of the
SUMOylation-deficient mutant. These findings indicate that SUMOylation
protects phosducin from proteasomal degradation. SUMOylation of phosducin
decreased its ability to bind Gbetagamma. PhlP, a closely related
member of the phosducin family, was not a target for SUMOylation,
but its SUMOylation can be achieved by a single amino acid insertion
in the conserved N terminus of PhlP. Together, these findings show
that phosducin is a previously unrecognized target of SUMO modification
and that SUMOylation controls phosducin stability in cells as well
as its functional properties.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Klenk, C. and Humrich, J. and Quitterer, U. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/2417e46879d727e2368e02efe697e4676/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {df527b874eb734ae7d21fcd3926e8aa8},
intrahash = {417e46879d727e2368e02efe697e4676},
issn = {0021-9258 (Print) 0021-9258 (Linking)},
journal = {J Biol Chem},
keywords = {& Acid Amino Animals Binding Blotting, COS Carrier Cell Cercopithecus Consensus Culture Data Eye G-Protein-Coupled/metabolism GTP-Binding Homology, Humans Line Lysine/chemistry Molecular Motifs Mutation Myocardium/chemistry Nerve Phosphoproteins/chemistry/genetics/isolation Phosphorylation Precipitin Protein Protein/genetics/*metabolism Proteins Proteins/chemistry/genetics/isolation Proteins/chemistry/metabolism Rats Recombinant Regulators Retina/chemistry Rhodopsin/metabolism SUMO-1 Sequence Sites Subunits/metabolism Techniques Tests Tissue Weight Western aethiops beta gamma purification/*metabolism Receptor},
month = {Mar 31},
note = {Klenk, Christoph Humrich, Jan Quitterer, Ursula Lohse, Martin J Comparative
Study Research Support, Non-U.S. Gov't United States The Journal
of biological chemistry J Biol Chem. 2006 Mar 31;281(13):8357-64.
Epub 2006 Jan 18.},
number = 13,
pages = {8357-64},
shorttitle = {SUMO-1 controls the protein stability and the biological function
of phosducin},
timestamp = {2010-12-14T18:22:02.000+0100},
title = {SUMO-1 controls the protein stability and the biological function
of phosducin},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16421094},
volume = 281,
year = 2006
}