NXP-2 association with SUMO-2 depends on lysines required for transcriptional repression
- Adam Rosendorff*,
- Shuhei Sakakibara*,
- Sixin Lu*,
- Elliott Kieff*,†,
- Yan Xuan‡,
- Alessandra DiBacco‡,
- Yujiang Shi‡,
- Yang Shi‡, and
- Grace Gill‡
- *Department of Medicine and Department of Microbiology and Molecular Genetics, Channing Laboratory, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115; and
- ‡Department of Pathology, Harvard Medical School, Boston, MA 02115
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Contributed by Elliott Kieff, February 8, 2006
Abstract
Small ubiquitin-like modifier (SUMO) modification of transcription factors is generally associated with repression. Reverse genetic analysis of SUMO-1, and -2 conserved residues emphasized the importance of dual charge reversals in abrogating the critical role of SUMO-2 K33, K35, and K42 in repression. GST-SUMO-2-affinity chromatography followed by liquid chromatography (LC)-MS analysis identified proteins that appeared to bind preferentially to WT SUMO-2 versus SUMO-2 K33E and K35E. LSD1, NXP-2, KIAA0809 (ARIP4), SAE2, RanGAP1, PELP1, and SETDB1 bound to SUMO-2 and not to SUMO-2 K33E, K42E, or K35E and K42E. Although LSD1 is a histone lysine demethylase, and histone H3K4 was demethylated at a SUMO-2-repressed promoter, neither overexpression of a dominant-negative LSD1 nor LSD1 depletion with RNA interference affected SUMO-2-mediated repression, indicating that LSD1 is not essential for repression, in this context. When tethered to a promoter by fusion to Gal4, NXP-2 repressed transcription, consistent with a role for NXP-2 in SUMO-mediated repression. SUMO-2-associated proteins identified in this study may contribute to SUMO-dependent regulation of transcription or other processes.
Footnotes
- †To whom correspondence should be addressed at: Channing Laboratory, 181 Longwood Avenue, Boston, MA 02114. E-mail: ekieff{at}rics.bwh.harvard.edu
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Author contributions: A.R., S.S., E.K., A.D., and G.G. designed research; A.R., S.S., S.L., Y.X., and A.D. performed research; Yujiang Shi and Yang Shi contributed new reagents/analytic tools; A.R., S.S., S.L., E.K., Y.X., A.D., and G.G. analyzed data; and A.R., S.S., E.K., and G.G. wrote the paper.
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Conflict of interest statement: No conflicts declared.
- Abbreviations:
- SUMO,
- small ubiquitin-like modifier.
Abbreviation:
- © 2006 by The National Academy of Sciences of the USA
