Proteomics

Dataset Information

2

The SUMO protease SENP6 regulates the constitutive centromere-associated network by group


ABSTRACT: Project 1: Identification of SUMOylated proteins which are regulated by the SUMO protease SENP6. SENP6 knockdown was performed by two independent shRNAs and SUMOylated proteins were purified from U2OS expressing His10-tagged SUMO2 by means of Ni-NTA pulldown. Purified SUMOylated proteins were identified and quantified by label-free quantification. Project 2: Dynamics of chromatin-associated proteins influenced by SENP6. After siRNA-mediated knockdown of SENP6, chromatin fractions of U2OS cells were isolated and abundances of proteins were quantified. Project 3: Analysis of in vitro SUMOylated CENP-T for the presence of SUMOylated SUMO peptides. For this aim purified CENP-T-HA was in vitro SUMOylated with a SUMO Q87R mutant to reduce the length of the tryptic peptide and therefore to be able to identified sit-specific SUMOylation.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo sapiens  

TISSUE(S): Cell Culture

DISEASE(S): Not Available

SUBMITTER: Frauke Liebelt  

LAB HEAD: Alfred C.O. Vertegaal

PROVIDER: PXD011963 | Pride | 2019-09-13

REPOSITORIES: Pride

Dataset's files

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Project 1 andromeda.zip Other
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Project 2 andromeda.zip Other
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Publications

The poly-SUMO2/3 protease SENP6 enables assembly of the constitutive centromere-associated network by group deSUMOylation.

Liebelt Frauke F   Jansen Nicolette S NS   Kumar Sumit S   Gracheva Ekaterina E   Claessens Laura A LA   Verlaan-de Vries Matty M   Willemstein Edwin E   Vertegaal Alfred C O ACO  

Nature communications 20190904 1


In contrast to our extensive knowledge on ubiquitin polymer signaling, we are severely limited in our understanding of poly-SUMO signaling. We set out to identify substrates conjugated to SUMO polymers, using knockdown of the poly-SUMO2/3 protease SENP6. We identify over 180 SENP6 regulated proteins that represent highly interconnected functional groups of proteins including the constitutive centromere-associated network (CCAN), the CENP-A loading factors Mis18BP1 and Mis18A and DNA damage respo  ...[more]

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