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Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII
EcoRII restriction endonuclease is specific for the 5′-CCWGG sequence (W stands for A or T); however, it shows no activity on a single recognition site. To activate cleavage it requires binding of an additional target site as an allosteric effector. EcoRII dimer consists of three structural units: a...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2770665/ https://www.ncbi.nlm.nih.gov/pubmed/19729506 http://dx.doi.org/10.1093/nar/gkp699 |
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author | Golovenko, Dmitrij Manakova, Elena Tamulaitiene, Giedre Grazulis, Saulius Siksnys, Virginijus |
author_facet | Golovenko, Dmitrij Manakova, Elena Tamulaitiene, Giedre Grazulis, Saulius Siksnys, Virginijus |
author_sort | Golovenko, Dmitrij |
collection | PubMed |
description | EcoRII restriction endonuclease is specific for the 5′-CCWGG sequence (W stands for A or T); however, it shows no activity on a single recognition site. To activate cleavage it requires binding of an additional target site as an allosteric effector. EcoRII dimer consists of three structural units: a central catalytic core, made from two copies of the C-terminal domain (EcoRII-C), and two N-terminal effector DNA binding domains (EcoRII-N). Here, we report DNA-bound EcoRII-N and EcoRII-C structures, which show that EcoRII combines two radically different structural mechanisms to interact with the effector and substrate DNA. The catalytic EcoRII-C dimer flips out the central T:A base pair and makes symmetric interactions with the CC:GG half-sites. The EcoRII-N effector domain monomer binds to the target site asymmetrically in a single defined orientation which is determined by specific hydrogen bonding and van der Waals interactions with the central T:A pair in the major groove. The EcoRII-N mode of the target site recognition is shared by the large class of higher plant transcription factors of the B3 superfamily. |
format | Text |
id | pubmed-2770665 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-27706652009-10-30 Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII Golovenko, Dmitrij Manakova, Elena Tamulaitiene, Giedre Grazulis, Saulius Siksnys, Virginijus Nucleic Acids Res Structural Biology EcoRII restriction endonuclease is specific for the 5′-CCWGG sequence (W stands for A or T); however, it shows no activity on a single recognition site. To activate cleavage it requires binding of an additional target site as an allosteric effector. EcoRII dimer consists of three structural units: a central catalytic core, made from two copies of the C-terminal domain (EcoRII-C), and two N-terminal effector DNA binding domains (EcoRII-N). Here, we report DNA-bound EcoRII-N and EcoRII-C structures, which show that EcoRII combines two radically different structural mechanisms to interact with the effector and substrate DNA. The catalytic EcoRII-C dimer flips out the central T:A base pair and makes symmetric interactions with the CC:GG half-sites. The EcoRII-N effector domain monomer binds to the target site asymmetrically in a single defined orientation which is determined by specific hydrogen bonding and van der Waals interactions with the central T:A pair in the major groove. The EcoRII-N mode of the target site recognition is shared by the large class of higher plant transcription factors of the B3 superfamily. Oxford University Press 2009-10 2009-09-03 /pmc/articles/PMC2770665/ /pubmed/19729506 http://dx.doi.org/10.1093/nar/gkp699 Text en © The Author(s) 2009. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Structural Biology Golovenko, Dmitrij Manakova, Elena Tamulaitiene, Giedre Grazulis, Saulius Siksnys, Virginijus Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title | Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title_full | Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title_fullStr | Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title_full_unstemmed | Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title_short | Structural mechanisms for the 5′-CCWGG sequence recognition by the N- and C-terminal domains of EcoRII |
title_sort | structural mechanisms for the 5′-ccwgg sequence recognition by the n- and c-terminal domains of ecorii |
topic | Structural Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2770665/ https://www.ncbi.nlm.nih.gov/pubmed/19729506 http://dx.doi.org/10.1093/nar/gkp699 |
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