Reactions of Type II Restriction Endonucleases with 8-Base Pair Recognition Sites

Type II restriction endonucleases usually recognize 4–6-base pair (bp) sites on DNA and cleave each site in a separate reaction. A few type II endonucleases have 8-bp recognition sites, but these seem unsuited for restriction, since their sites are rare on most DNA. Moreover, only one endonuclease...

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Veröffentlicht in:The Journal of biological chemistry 1999-12, Vol.274 (51), p.36379-36386
Hauptverfasser: Bilcock, D T, Daniels, L E, Bath, A J, Halford, S E
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container_end_page 36386
container_issue 51
container_start_page 36379
container_title The Journal of biological chemistry
container_volume 274
creator Bilcock, D T
Daniels, L E
Bath, A J
Halford, S E
description Type II restriction endonucleases usually recognize 4–6-base pair (bp) sites on DNA and cleave each site in a separate reaction. A few type II endonucleases have 8-bp recognition sites, but these seem unsuited for restriction, since their sites are rare on most DNA. Moreover, only one endonuclease that recognizes a target containing 8 bp has been examined to date, and this enzyme, Sfi I, needs two copies of this site for its DNA cleavage reaction. In this study, several endonucleases with 8-bp sites were tested on plasmids that have either one or two copies of the relevant sequence to determine if they also need two sites. Sgf I, Srf I, Fse I, Pac I, Pme I, Sse 8781I, and Sda I all acted through equal and independent reactions at each site. Asc I cleaved the DNA with one site at the same rate as that with two sites but acted processively on the latter. In contrast, Sgr AI showed a marked preference for the plasmid with two sites and cleaved both sites on this DNA in a concerted manner, like Sfi I. Endonucleases that require two copies of an 8-bp sequence may be widespread in nature, where, despite this seemingly inappropriate requirement, they may function in DNA restriction.
doi_str_mv 10.1074/jbc.274.51.36379
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subjects Base Pairing
Binding Sites - genetics
deoxyribonuclease FseI
deoxyribonuclease PacI
deoxyribonuclease PmeI
deoxyribonuclease SfiI
deoxyribonuclease SgfI
deoxyribonuclease SrfI
Deoxyribonucleases, Type II Site-Specific - chemistry
Deoxyribonucleases, Type II Site-Specific - genetics
Deoxyribonucleases, Type II Site-Specific - metabolism
DNA - genetics
DNA - metabolism
Plasmids
Sequence Analysis
Substrate Specificity
title Reactions of Type II Restriction Endonucleases with 8-Base Pair Recognition Sites
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