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Published online 26 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2287-2297
© 2004 Oxford University Press

SCFhFBH1 can act as helicase and E3 ubiquitin ligase

Jeong-Hoon Kim, Jaehoon Kim, Do-Hyung Kim, Gi-Hyuck Ryu, Sung-Ho Bae1 and Yeon-Soo Seo*

National Creative Research Initiative Center for Cell Cycle Control, Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejoen, 305-701, Korea and 1 Department of Biological Sciences, Inha University, 253, Yonghyun-Dong, Nam-Ku, Inchon, 402-751, Korea

*To whom correspondence should be addressed. Tel: +82 42 869 2637; Fax: +82 42 869 2610; Email: yeonsooseo{at}kaist.ac.kr

Received September 23, 2003; Revised October 24, 2003; Accepted November 13, 2003

In our previous study, we found that a human F-box DNA helicase, named hFBH1, interacted with SKP1 to form an SCF (SKP1–Cul1–F-box protein) complex together with CUL1 and ROC1 in an F-box-dependent manner. The complex immunoprecipitated from crude cell extracts catalyzed polyubiquitin formation in the presence of the ubiquitin-activating and ubiquitin-conjugating enzymes, E1 and E2, respectively. In this report, we characterized the enzymatic properties of the recombinant SCFhFBH1 complex purified from insect cells expressing hFBH1, SKP1, CUL1 and ROC1. The SCFhFBH1 complex was isolated as a single tight complex that retained DNA helicase, DNA-dependent ATPase and E3 ubiquitin ligase activities. The helicase and ATPase activities residing in the SCFhFBH1 complex were indistinguishable from those of the hFBH1 protein alone. Moreover, the ubiquitin ligase activity of the SCFhFBH1 complex was hardly affected by single-stranded or double-stranded DNA. The multiple activities present in this complex act independently of each other, suggesting that the SCFhFBH1 complex can catalyze a ubiquitination reaction while acting as a DNA helicase or translocating along DNA. The potential roles of the SCFhFBH1 complex in DNA metabolism based upon the enzymatic activities associated with this complex are discussed.


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