Sumoylation of Hes6 regulates protein degradation and Hes1-mediated transcription

Jiwon Lee, Sung Kook Chun, Gi Hoon Son, Kyungjin Kim

Research output: Contribution to journalArticle

Abstract

Background: Hes6 is a transcriptional regulator that induces transcriptional activation by binding to transcription repressor Hes1 and suppressing its activity. Hes6 is controlled by the ubiquitin-proteosome-mediated degradation system. Here we investigated the sumoylation of Hes6 and its functional role in its rhythmic expression. Methods: Hes6, SUMO, and ubiquitin were transfected into HeLa cells and the expression pattern was observed by Western blot and immunoprecipitation. To confirm the effect of sumoylation on the rhythmic expression of Hes6, we generated mouse Hes6 promoter-driven GFP-Hes6 fusion constructs and expressed these constructs in NIH 3T3 cells. Results: Overexpression of SUMO led to sumoylation of Hes6 at both lysine 27 and 30. Protein stability of Hes6 was decreased by sumoylation. Moreover, expression of a Hes6 sumoylation-defective mutant, the 2KR (K27/30R) mutant, or co-expression of SUMO protease SUSP1 with native Hes6, strongly reduced ubiquitination. In addition, sumoylation was associated with both the rhythmic expression and transcriptional regulation of Hes6. Wild type Hes6 showed oscillatory expression with about 2-hour periodicity, whereas the 2KR mutant displayed a longer period. Furthermore, sumoylation of Hes6 derepressed Hes1-induced transcriptional repression. Conclusion: Hes6 sumoylation plays an important role in the regulation of its stability and Hes1-mediated transcription. These results suggest that sumoylation may be crucial for rhythmic expression of Hes6 and downstream target genes.

Original languageEnglish
Pages (from-to)381-388
Number of pages8
JournalEndocrinology and Metabolism
Volume30
Issue number3
DOIs
Publication statusPublished - 2015

Fingerprint

Sumoylation
Proteolysis
Ubiquitin
NIH 3T3 Cells
Protein Stability
Ubiquitination
Periodicity
HeLa Cells
Immunoprecipitation
Transcriptional Activation
Lysine
Peptide Hydrolases
Western Blotting

Keywords

  • Degradation
  • Hes6
  • Rhythmic expression
  • Sumoylation
  • Ubiquitination

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Sumoylation of Hes6 regulates protein degradation and Hes1-mediated transcription. / Lee, Jiwon; Chun, Sung Kook; Son, Gi Hoon; Kim, Kyungjin.

In: Endocrinology and Metabolism, Vol. 30, No. 3, 2015, p. 381-388.

Research output: Contribution to journalArticle

Lee, Jiwon ; Chun, Sung Kook ; Son, Gi Hoon ; Kim, Kyungjin. / Sumoylation of Hes6 regulates protein degradation and Hes1-mediated transcription. In: Endocrinology and Metabolism. 2015 ; Vol. 30, No. 3. pp. 381-388.
@article{058a3d0fd9b24cd584610f4144b7f74f,
title = "Sumoylation of Hes6 regulates protein degradation and Hes1-mediated transcription",
abstract = "Background: Hes6 is a transcriptional regulator that induces transcriptional activation by binding to transcription repressor Hes1 and suppressing its activity. Hes6 is controlled by the ubiquitin-proteosome-mediated degradation system. Here we investigated the sumoylation of Hes6 and its functional role in its rhythmic expression. Methods: Hes6, SUMO, and ubiquitin were transfected into HeLa cells and the expression pattern was observed by Western blot and immunoprecipitation. To confirm the effect of sumoylation on the rhythmic expression of Hes6, we generated mouse Hes6 promoter-driven GFP-Hes6 fusion constructs and expressed these constructs in NIH 3T3 cells. Results: Overexpression of SUMO led to sumoylation of Hes6 at both lysine 27 and 30. Protein stability of Hes6 was decreased by sumoylation. Moreover, expression of a Hes6 sumoylation-defective mutant, the 2KR (K27/30R) mutant, or co-expression of SUMO protease SUSP1 with native Hes6, strongly reduced ubiquitination. In addition, sumoylation was associated with both the rhythmic expression and transcriptional regulation of Hes6. Wild type Hes6 showed oscillatory expression with about 2-hour periodicity, whereas the 2KR mutant displayed a longer period. Furthermore, sumoylation of Hes6 derepressed Hes1-induced transcriptional repression. Conclusion: Hes6 sumoylation plays an important role in the regulation of its stability and Hes1-mediated transcription. These results suggest that sumoylation may be crucial for rhythmic expression of Hes6 and downstream target genes.",
keywords = "Degradation, Hes6, Rhythmic expression, Sumoylation, Ubiquitination",
author = "Jiwon Lee and Chun, {Sung Kook} and Son, {Gi Hoon} and Kyungjin Kim",
year = "2015",
doi = "10.3803/EnM.2015.30.3.381",
language = "English",
volume = "30",
pages = "381--388",
journal = "Endocrinology and Metabolism",
issn = "2093-596X",
publisher = "Korean Endocrine Society",
number = "3",

}

TY - JOUR

T1 - Sumoylation of Hes6 regulates protein degradation and Hes1-mediated transcription

AU - Lee, Jiwon

AU - Chun, Sung Kook

AU - Son, Gi Hoon

AU - Kim, Kyungjin

PY - 2015

Y1 - 2015

N2 - Background: Hes6 is a transcriptional regulator that induces transcriptional activation by binding to transcription repressor Hes1 and suppressing its activity. Hes6 is controlled by the ubiquitin-proteosome-mediated degradation system. Here we investigated the sumoylation of Hes6 and its functional role in its rhythmic expression. Methods: Hes6, SUMO, and ubiquitin were transfected into HeLa cells and the expression pattern was observed by Western blot and immunoprecipitation. To confirm the effect of sumoylation on the rhythmic expression of Hes6, we generated mouse Hes6 promoter-driven GFP-Hes6 fusion constructs and expressed these constructs in NIH 3T3 cells. Results: Overexpression of SUMO led to sumoylation of Hes6 at both lysine 27 and 30. Protein stability of Hes6 was decreased by sumoylation. Moreover, expression of a Hes6 sumoylation-defective mutant, the 2KR (K27/30R) mutant, or co-expression of SUMO protease SUSP1 with native Hes6, strongly reduced ubiquitination. In addition, sumoylation was associated with both the rhythmic expression and transcriptional regulation of Hes6. Wild type Hes6 showed oscillatory expression with about 2-hour periodicity, whereas the 2KR mutant displayed a longer period. Furthermore, sumoylation of Hes6 derepressed Hes1-induced transcriptional repression. Conclusion: Hes6 sumoylation plays an important role in the regulation of its stability and Hes1-mediated transcription. These results suggest that sumoylation may be crucial for rhythmic expression of Hes6 and downstream target genes.

AB - Background: Hes6 is a transcriptional regulator that induces transcriptional activation by binding to transcription repressor Hes1 and suppressing its activity. Hes6 is controlled by the ubiquitin-proteosome-mediated degradation system. Here we investigated the sumoylation of Hes6 and its functional role in its rhythmic expression. Methods: Hes6, SUMO, and ubiquitin were transfected into HeLa cells and the expression pattern was observed by Western blot and immunoprecipitation. To confirm the effect of sumoylation on the rhythmic expression of Hes6, we generated mouse Hes6 promoter-driven GFP-Hes6 fusion constructs and expressed these constructs in NIH 3T3 cells. Results: Overexpression of SUMO led to sumoylation of Hes6 at both lysine 27 and 30. Protein stability of Hes6 was decreased by sumoylation. Moreover, expression of a Hes6 sumoylation-defective mutant, the 2KR (K27/30R) mutant, or co-expression of SUMO protease SUSP1 with native Hes6, strongly reduced ubiquitination. In addition, sumoylation was associated with both the rhythmic expression and transcriptional regulation of Hes6. Wild type Hes6 showed oscillatory expression with about 2-hour periodicity, whereas the 2KR mutant displayed a longer period. Furthermore, sumoylation of Hes6 derepressed Hes1-induced transcriptional repression. Conclusion: Hes6 sumoylation plays an important role in the regulation of its stability and Hes1-mediated transcription. These results suggest that sumoylation may be crucial for rhythmic expression of Hes6 and downstream target genes.

KW - Degradation

KW - Hes6

KW - Rhythmic expression

KW - Sumoylation

KW - Ubiquitination

UR - http://www.scopus.com/inward/record.url?scp=84955500584&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84955500584&partnerID=8YFLogxK

U2 - 10.3803/EnM.2015.30.3.381

DO - 10.3803/EnM.2015.30.3.381

M3 - Article

VL - 30

SP - 381

EP - 388

JO - Endocrinology and Metabolism

JF - Endocrinology and Metabolism

SN - 2093-596X

IS - 3

ER -