TY - JOUR
T1 - A proteome-wide approach identifies sumoylated substrate proteins in yeast
AU - Panse, Vikram Govind
AU - Hardeland, Ulrike
AU - Werner, Thilo
AU - Kuster, Bernhard
AU - Hurt, Ed
PY - 2004/10/1
Y1 - 2004/10/1
N2 - The ubiquitin-related protein SUMO-1 is covalently attached to proteins by SUMO-1 ligases. We have performed a proteome-wide analysis of sumoylated substrate proteins in yeast. Employing the powerful affinity purification of Protein A-Smt3 (Smt3 is the yeast homologue of SUMO-1) from yeast lysates in combination with tandem liquid chromatography mass spectrometry, we have isolated potential Smt3-carrying substrate proteins involved in DNA replication and repair, chromatin remodeling, transcription activation, Pol-I, Pol-II, and Pol-III transcription, 5′ pre-mRNA capping, 3′ pre-mRNA processing, proteasome function, and tubulin folding. Employing tandem affinity purifications or a rapid biochemical assay referred to as "SUMO fingerprint," we showed that several subunits of RNA polymerases I, II, and III, members of the transcription repression and chromatin remodeling machineries previously not known to be sumoylated, are modified by SUMO-1. Thus, the identification of a broad range of SUMO-1 substrate proteins is expected to lead to further insight into the regulatory aspects of sumoylation.
AB - The ubiquitin-related protein SUMO-1 is covalently attached to proteins by SUMO-1 ligases. We have performed a proteome-wide analysis of sumoylated substrate proteins in yeast. Employing the powerful affinity purification of Protein A-Smt3 (Smt3 is the yeast homologue of SUMO-1) from yeast lysates in combination with tandem liquid chromatography mass spectrometry, we have isolated potential Smt3-carrying substrate proteins involved in DNA replication and repair, chromatin remodeling, transcription activation, Pol-I, Pol-II, and Pol-III transcription, 5′ pre-mRNA capping, 3′ pre-mRNA processing, proteasome function, and tubulin folding. Employing tandem affinity purifications or a rapid biochemical assay referred to as "SUMO fingerprint," we showed that several subunits of RNA polymerases I, II, and III, members of the transcription repression and chromatin remodeling machineries previously not known to be sumoylated, are modified by SUMO-1. Thus, the identification of a broad range of SUMO-1 substrate proteins is expected to lead to further insight into the regulatory aspects of sumoylation.
UR - http://www.scopus.com/inward/record.url?scp=4744360999&partnerID=8YFLogxK
U2 - 10.1074/jbc.M407950200
DO - 10.1074/jbc.M407950200
M3 - Article
C2 - 15292183
AN - SCOPUS:4744360999
SN - 0021-9258
VL - 279
SP - 41346
EP - 41351
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 40
ER -