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Title
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Crystallization and preliminary X-ray characterization of the eukaryotic replication terminator Reb1-Ter DNA complex.
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Authors
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R.Jaiswal,
S.K.Singh,
D.Bastia,
C.R.Escalante.
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Ref.
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Acta Crystallogr F Struct Biol Commun, 2015,
71,
414-418.
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PubMed id
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Abstract
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The Reb1 protein from Schizosaccharomyces pombe is a member of a family of
proteins that control programmed replication termination and/or transcription
termination in eukaryotic cells. These events occur at naturally occurring
replication fork barriers (RFBs), where Reb1 binds to termination (Ter) DNA
sites and coordinates the polar arrest of replication forks and transcription
approaching in opposite directions. The Reb1 DNA-binding and
replication-termination domain was expressed in Escherichia coli, purified and
crystallized in complex with a 26-mer DNA Ter site. Batch crystallization under
oil was required to produce crystals of good quality for data collection.
Crystals grew in space group P2₁, with unit-cell parameters a = 68.9, b =
162.9, c = 71.1 Å, β = 94.7°. The crystals diffracted to a resolution of
3.0 Å. The crystals were mosaic and required two or three cycles of
annealing. This study is the first to yield structural information about this
important family of proteins and will provide insights into the mechanism of
replication and transcription termination.
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