Assembly of a polymeric chain of SUMO1 on human topoisomerase I in vitro

Meiluen Yang, Chia Tse Hsu, Chun Yuan Ting, Leroy-Fong Liu, Jaulang Hwang

研究成果: 雜誌貢獻文章同行評審

53 引文 斯高帕斯(Scopus)

摘要

Human (h) DNA topoisomerase I has been identified as a major SUMO1 target in camptothecin-treated cells. In response to TOP1-mediated DNA damage induced by camptothecin, multiple SUMO1 molecules are conjugated to the N-terminal domain of a single TOP1 molecule. To investigate the molecular mechanism of SUMO1 conjugation to TOP1, an in vitro system using purified SAE1/2, Ubc9, SUMO1, and TOP1 peptides was developed. Consistent with results from in vivo studies, multiple SUMO1 molecules were found to be conjugated to the N-terminal domain of a single TOP1 molecule. Systematic analysis has identified a single major SUMO1 conjugation site located between amino acid residues 110 and 125 that contains a single lysine residue at 117 (Lys-117). Using a short peptide spanning this region, we showed that a poly-SUMO1 chain was assembled in this peptide at Lys-117. Interestingly, a Ubc9-poly-SUMO1 intermediate had accumulated to a high level when the sumoylation assay was performed in the absence of hTOP1 substrate, suggesting a possibility that the poly-SUMO1 chain is formed on Ubc9 first and then transferred en bloc onto hTOP1. This is the first definitive demonstration of the assembly of a poly-SUMO1 chain on protein substrate. These results offer new insight into hTOP1 polysumoylation in response to TOP1-mediated DNA damage and may have general implications in protein polysumoylation.

原文英語
頁(從 - 到)8264-8274
頁數11
期刊Journal of Biological Chemistry
281
發行號12
DOIs
出版狀態已發佈 - 三月 24 2006
對外發佈Yes

ASJC Scopus subject areas

  • Biochemistry

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