Systematic identification and analysis of mammalian small ubiquitin-like modifier substrates

Christian B. Gocke, Hongtao Yu, Jungseog Kang

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

Small ubiquitin-like modifier (SUMO) regulates diverse cellular processes through its reversible, covalent attachment to target proteins. Many SUMO substrates are involved in transcription and chromatin structure. Sumoylation appears to regulate the functions of target proteins by changing their subcellular localization, increasing their stability, and/or mediating their binding to other proteins. Using an in vitro expression cloning approach, we have identified 40 human SUMO1 substrates. The spectrum of human SUMO1 substrates identified in our screen suggests general roles of sumoylation in transcription, chromosome structure, and RNA processing. We have validated the sumoylation of 24 substrates in living cells. Analysis of this panel of SUMO substrates leads to the following observations. 1) Sumoylation is more efficient in vitro than in living cells. Polysumoylation occurs on several substrates in vitro. 2) SUMO isopeptidases have little substrate specificity. 3) The SUMO ligases, PIAS1 and PIASxβ, have broader substrate specificities than does PIASy. 4) Although SUMO1 and SUMO2 are equally efficiently conjugated to a given substrate in vitro, SUMO1 conjugation is more efficient in vivo. 5) Most SUMO substrates localize to the nucleus, and sumoylation does not generally affect their subcellular localization. Therefore, sumoylation appears to regulate the functions of its substrates through multiple, context-dependent mechanisms.

Original languageEnglish (US)
Pages (from-to)5004-5012
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number6
DOIs
StatePublished - Feb 11 2005

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Sumoylation
Ubiquitin
Substrates
Substrate Specificity
Chromosome Structures
Proteins
Transcription
Ligases
Chromatin
Organism Cloning
Cells
RNA
Cloning
In Vitro Techniques
Chromosomes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Systematic identification and analysis of mammalian small ubiquitin-like modifier substrates. / Gocke, Christian B.; Yu, Hongtao; Kang, Jungseog.

In: Journal of Biological Chemistry, Vol. 280, No. 6, 11.02.2005, p. 5004-5012.

Research output: Contribution to journalArticle

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