Crystal structure of the human Hsmar1-derived transposase domain in the DNA repair enzyme Metnase.

Kristie D. Goodwin, Hongzhen He, Tsuyoshi Imasaki, Suk-Hee Lee, Millie Georgiadis

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Although the human genome is littered with sequences derived from the Hsmar1 transposon, the only intact Hsmar1 transposase gene exists within a chimeric SET-transposase fusion protein referred to as Metnase or SETMAR. Metnase retains many of the transposase activities including terminal inverted repeat (TIR) specific DNA-binding activity, DNA cleavage activity, albeit uncoupled from TIR-specific binding, and the ability to form a synaptic complex. However, Metnase has evolved as a DNA repair protein that is specifically involved in nonhomologous end joining. Here, we present two crystal structures of the transposase catalytic domain of Metnase revealing a dimeric enzyme with unusual active site plasticity that may be involved in modulating metal binding.We show through characterization of a dimerization mutant, F460K, that the dimeric form of the enzyme is required for itsDNAcleavage, DNA-binding, and nonhomologous end joining activities. Of significance is the conservation of F460 along with residues that we propose may be involved in the modulation of metal binding in both the predicted ancestral Hsmar1 transposase sequence as well as in the modern enzyme. The Metnase transposase has been remarkably conserved through evolution; however, there is a clustering of substitutions located in alpha helices 4 and 5 within the putative DNAbinding site, consistent with loss of transposition specific DNA cleavage activity and acquisition of DNA repair specific cleavage activity.

Original languageEnglish
Pages (from-to)5705-5713
Number of pages9
JournalBiochemistry
Volume49
Issue number27
DOIs
StatePublished - Jul 13 2010

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DNA Repair Enzymes
Transposases
Crystal structure
DNA
DNA Cleavage
Terminal Repeat Sequences
Joining
DNA Repair
Catalytic Domain
Repair
Enzymes
DNA End-Joining Repair
Genes
Metals
Dimerization
Human Genome
Plasticity
Cluster Analysis
Conservation
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Crystal structure of the human Hsmar1-derived transposase domain in the DNA repair enzyme Metnase. / Goodwin, Kristie D.; He, Hongzhen; Imasaki, Tsuyoshi; Lee, Suk-Hee; Georgiadis, Millie.

In: Biochemistry, Vol. 49, No. 27, 13.07.2010, p. 5705-5713.

Research output: Contribution to journalArticle

Goodwin, Kristie D. ; He, Hongzhen ; Imasaki, Tsuyoshi ; Lee, Suk-Hee ; Georgiadis, Millie. / Crystal structure of the human Hsmar1-derived transposase domain in the DNA repair enzyme Metnase. In: Biochemistry. 2010 ; Vol. 49, No. 27. pp. 5705-5713.
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