Biochemical characterization of metnases endonuclease activity and its role in NHEJ repair

Brian D. Beck; Sung Sook Lee; Elizabeth Williamson; Robert A. Hromas; Suk Hee Lee

doi:10.1021/bi200333k

Biochemical characterization of metnases endonuclease activity and its role in NHEJ repair

Brian D. Beck, Sung Sook Lee, Elizabeth Williamson, Robert A. Hromas, Suk Hee Lee

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

Abstract

Metnase (SETMAR) is a SET-transposase fusion protein that promotes nonhomologous end joining (NHEJ) repair in humans. Although both SET and the transposase domains were necessary for its function in DSB repair, it is not clear what specific role Metnase plays in the NHEJ. In this study, we show that Metnase possesses a unique endonuclease activity that preferentially acts on ssDNA and ssDNA-overhang of a partial duplex DNA. Cell extracts lacking Metnase poorly supported DNA end joining, and addition of wt-Metnase to cell extracts lacking Metnase markedly stimulated DNA end joining, while a mutant (D483A) lacking endonuclease activity did not. Given that Metnase overexpression enhanced DNA end processing in vitro, our finding suggests a role for Metnases endonuclease activity in promoting the joining of noncompatible ends.

Original language	English (US)
Pages (from-to)	4360-4370
Number of pages	11
Journal	Biochemistry
Volume	50
Issue number	20
DOIs	https://doi.org/10.1021/bi200333k
State	Published - May 24 2011
Externally published	Yes

ASJC Scopus subject areas

Biochemistry

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10.1021/bi200333k

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title = "Biochemical characterization of metnases endonuclease activity and its role in NHEJ repair",

abstract = "Metnase (SETMAR) is a SET-transposase fusion protein that promotes nonhomologous end joining (NHEJ) repair in humans. Although both SET and the transposase domains were necessary for its function in DSB repair, it is not clear what specific role Metnase plays in the NHEJ. In this study, we show that Metnase possesses a unique endonuclease activity that preferentially acts on ssDNA and ssDNA-overhang of a partial duplex DNA. Cell extracts lacking Metnase poorly supported DNA end joining, and addition of wt-Metnase to cell extracts lacking Metnase markedly stimulated DNA end joining, while a mutant (D483A) lacking endonuclease activity did not. Given that Metnase overexpression enhanced DNA end processing in vitro, our finding suggests a role for Metnases endonuclease activity in promoting the joining of noncompatible ends.",

author = "Beck, {Brian D.} and Lee, {Sung Sook} and Elizabeth Williamson and Hromas, {Robert A.} and Lee, {Suk Hee}",

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T1 - Biochemical characterization of metnases endonuclease activity and its role in NHEJ repair

AU - Beck, Brian D.

AU - Lee, Sung Sook

AU - Williamson, Elizabeth

AU - Hromas, Robert A.

AU - Lee, Suk Hee

PY - 2011/5/24

Y1 - 2011/5/24

N2 - Metnase (SETMAR) is a SET-transposase fusion protein that promotes nonhomologous end joining (NHEJ) repair in humans. Although both SET and the transposase domains were necessary for its function in DSB repair, it is not clear what specific role Metnase plays in the NHEJ. In this study, we show that Metnase possesses a unique endonuclease activity that preferentially acts on ssDNA and ssDNA-overhang of a partial duplex DNA. Cell extracts lacking Metnase poorly supported DNA end joining, and addition of wt-Metnase to cell extracts lacking Metnase markedly stimulated DNA end joining, while a mutant (D483A) lacking endonuclease activity did not. Given that Metnase overexpression enhanced DNA end processing in vitro, our finding suggests a role for Metnases endonuclease activity in promoting the joining of noncompatible ends.

AB - Metnase (SETMAR) is a SET-transposase fusion protein that promotes nonhomologous end joining (NHEJ) repair in humans. Although both SET and the transposase domains were necessary for its function in DSB repair, it is not clear what specific role Metnase plays in the NHEJ. In this study, we show that Metnase possesses a unique endonuclease activity that preferentially acts on ssDNA and ssDNA-overhang of a partial duplex DNA. Cell extracts lacking Metnase poorly supported DNA end joining, and addition of wt-Metnase to cell extracts lacking Metnase markedly stimulated DNA end joining, while a mutant (D483A) lacking endonuclease activity did not. Given that Metnase overexpression enhanced DNA end processing in vitro, our finding suggests a role for Metnases endonuclease activity in promoting the joining of noncompatible ends.

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Biochemical characterization of metnases endonuclease activity and its role in NHEJ repair

Abstract

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