TY - JOUR
T1 - Subunit-selective proteasome activity profiling uncovers uncoupled proteasome subunit activities during bacterial infections
AU - Misas-villamil, Johana C.
AU - Van Der Burgh, Aranka M.
AU - Grosse-holz, Friederike
AU - Bach-pages, Marcel
AU - Kovács, Judit
AU - Kaschani, Farnusch
AU - Schilasky, Sören
AU - Emon, Asif E.K.
AU - Ruben, Mark
AU - Kaiser, Markus
AU - Overkleeft, Hermen S.
AU - van der Hoorn, Renier A.L.
PY - 2017
Y1 - 2017
N2 - The proteasome is a nuclear-cytoplasmic proteolytic complex involved in nearly all regulatory pathways in plant cells. The three different catalytic activities of the proteasome can have different functions, but tools to monitor and control these subunits selectively are not yet available in plant science. Here, we introduce subunit-selective inhibitors and dual-color fluorescent activity-based probes for studying two of the three active catalytic subunits of the plant proteasome. We validate these tools in two model plants and use this to study the proteasome during plant–microbe interactions. Our data reveal that Nicotiana benthamiana incorporates two different paralogs of each catalytic subunit into active proteasomes. Interestingly, both β1 and β5 activities are significantly increased upon infection with pathogenic Pseudomonas syringae pv. tomato DC3000 lacking hopQ1-1 [PtoDC3000(ΔhQ)] whilst the activity profile of the β1 subunit changes. Infection with wild-type PtoDC3000 causes proteasome activities that range from strongly induced β1 and β5 activities to strongly suppressed β5 activities, revealing that β1 and β5 activities can be uncoupled during bacterial infection. These selective probes and inhibitors are now available to the plant science community, and can be widely and easily applied to study the activity and role of the different catalytic subunits of the proteasome in different plant species.
AB - The proteasome is a nuclear-cytoplasmic proteolytic complex involved in nearly all regulatory pathways in plant cells. The three different catalytic activities of the proteasome can have different functions, but tools to monitor and control these subunits selectively are not yet available in plant science. Here, we introduce subunit-selective inhibitors and dual-color fluorescent activity-based probes for studying two of the three active catalytic subunits of the plant proteasome. We validate these tools in two model plants and use this to study the proteasome during plant–microbe interactions. Our data reveal that Nicotiana benthamiana incorporates two different paralogs of each catalytic subunit into active proteasomes. Interestingly, both β1 and β5 activities are significantly increased upon infection with pathogenic Pseudomonas syringae pv. tomato DC3000 lacking hopQ1-1 [PtoDC3000(ΔhQ)] whilst the activity profile of the β1 subunit changes. Infection with wild-type PtoDC3000 causes proteasome activities that range from strongly induced β1 and β5 activities to strongly suppressed β5 activities, revealing that β1 and β5 activities can be uncoupled during bacterial infection. These selective probes and inhibitors are now available to the plant science community, and can be widely and easily applied to study the activity and role of the different catalytic subunits of the proteasome in different plant species.
KW - activity-based protein profiling
KW - Arabidopsis thaliana
KW - catalytic subunit
KW - core protease
KW - Nicotiana benthamiana
KW - proteasome manipulation
KW - technical advance
U2 - 10.1111/tpj.13494
DO - 10.1111/tpj.13494
M3 - Article
SN - 0960-7412
VL - 90
SP - 418
EP - 430
JO - The Plant Journal
JF - The Plant Journal
IS - 2
ER -