Sumoylation of internally initiated Sp3 isoforms regulates transcriptional repression via a Trichostatin A-insensitive mechanism

Mary L. Spengler, Sarah B. Kennett, K. Scott Moorefield, Steven O. Simmons, Michael G. Brattain, Jonathan M. Horowitz

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Sp3 is a ubiquitously expressed member of the Sp family of transcription factors that encodes three proteins, Sp3, M1 and M2, with differing capacities to stimulate or repress transcription. As part of ongoing efforts to study the functions of Sp3 isoforms, we employed a yeast "two-hybrid" screen to identify Sp3-binding proteins. This screen resulted in the identification of Ubc9, a SUMO-1 conjugating enzyme, as an M2-binding protein, and consistent with these results sequence analyses identified consensus sumoylation motifs within several Sp family members. Western blots probed with anti-Sp3 detected a high molecular weight Sp3 isoform that is stabilized by a SUMO-1 hydrolase inhibitor, and this protein is also bound by anti-SUMO-1 antiserum. Transient transfection assays with epitope-tagged-SUMO-1 and GFP-SUMO-1 fusion proteins confirmed that Sp3, M1 and M2 proteins are sumoylated in vivo. Substitution of arginine for lysine at one putative site of sumoylation, lysine 551, blocked sumoylation of all Sp3 isoforms in vivo and led to a marginal increase in Sp3-mediated trans-activation in insect and mammalian cells. In contrast, introduction of this amino acid substitution within M1 converted it into a potent transcriptional trans-activator. We conclude that Sp3 isoforms are sumoylated in vivo and this post-translational modification plays an important role in the regulation of Sp3-mediated transcription.

Original languageEnglish (US)
Pages (from-to)153-166
Number of pages14
JournalCellular Signalling
Volume17
Issue number2
DOIs
StatePublished - Feb 1 2005

Fingerprint

trichostatin A
Sumoylation
Protein Isoforms
Lysine
Sp Transcription Factors
SUMO-1 Protein
Carrier Proteins
Proteins
Trans-Activators
Amino Acid Substitution
Post Translational Protein Processing
Transfection
Sequence Analysis
Insects
Arginine
Immune Sera
Epitopes
Yeasts
Molecular Weight
Western Blotting

Keywords

  • PML
  • Sp3
  • Sumoylation
  • Trans-activation
  • Trans-repression
  • Transcriptional regulation
  • Trichostatin A

ASJC Scopus subject areas

  • Cell Biology

Cite this

Spengler, M. L., Kennett, S. B., Moorefield, K. S., Simmons, S. O., Brattain, M. G., & Horowitz, J. M. (2005). Sumoylation of internally initiated Sp3 isoforms regulates transcriptional repression via a Trichostatin A-insensitive mechanism. Cellular Signalling, 17(2), 153-166. https://doi.org/10.1016/j.cellsig.2004.06.007

Sumoylation of internally initiated Sp3 isoforms regulates transcriptional repression via a Trichostatin A-insensitive mechanism. / Spengler, Mary L.; Kennett, Sarah B.; Moorefield, K. Scott; Simmons, Steven O.; Brattain, Michael G.; Horowitz, Jonathan M.

In: Cellular Signalling, Vol. 17, No. 2, 01.02.2005, p. 153-166.

Research output: Contribution to journalArticle

Spengler, Mary L. ; Kennett, Sarah B. ; Moorefield, K. Scott ; Simmons, Steven O. ; Brattain, Michael G. ; Horowitz, Jonathan M. / Sumoylation of internally initiated Sp3 isoforms regulates transcriptional repression via a Trichostatin A-insensitive mechanism. In: Cellular Signalling. 2005 ; Vol. 17, No. 2. pp. 153-166.
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