Structural basis for DNA recognition and loading into a viral packaging motor

Carina R. Buettner; Maria Chechik; Miguel Ortiz-Lombardia; Callum Smits; Ima-Obong Ebong; Victor Chechik; Gunnar Jeschke; Eric Dykeman; Stefano Benini; Carol V. Robinson; Juan C. Alonso; Alfred A. Antson

doi:10.1073/pnas.1110270109

Structural basis for DNA recognition and loading into a viral packaging motor

Carina R. Buettner, Maria Chechik, Miguel Ortiz-Lombardia, Callum Smits, Ima-Obong Ebong, Victor Chechik, Gunnar Jeschke, Eric Dykeman, Stefano Benini, Carol V. Robinson, Juan C. Alonso, Alfred A. Antson

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

Abstract

Genome packaging into preformed viral procapsids is driven by powerful molecular motors. The small terminase protein is essential for the initial recognition of viral DNA and regulates the motor's ATPase and nuclease activities during DNA translocation. The crystal structure of a full-length small terminase protein from the Siphoviridae bacteriophage SF6, comprising the N-terminal DNA binding, the oligomerization core, and the C-terminal beta-barrel domains, reveals a nine-subunit circular assembly in which the DNA-binding domains are arranged around the oligomerization core in a highly flexible manner. Mass spectrometry analysis and four further crystal structures show that, although the full-length protein exclusively forms nine-subunit assemblies, protein constructs missing the C-terminal beta-barrel form both nine-subunit and ten-subunit assemblies, indicating the importance of the C terminus for defining the oligomeric state. The mechanism by which a ring-shaped small terminase oligomer binds viral DNA has not previously been elucidated. Here, we probed binding in vitro by using EPR and surface plasmon resonance experiments, which indicated that interaction with DNA is mediated exclusively by the DNA-binding domains and suggested a nucleosome-like model in which DNA binds around the outside of the protein oligomer.

Original language	English
Pages (from-to)	811-816
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	109
Issue number	3
DOIs	https://doi.org/10.1073/pnas.1110270109
Publication status	Published - 17 Jan 2012

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10.1073/pnas.1110270109

http://www.pnas.org/content/109/3/811

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Buettner, C. R., Chechik, M., Ortiz-Lombardia, M., Smits, C., Ebong, I-O., Chechik, V., Jeschke, G., Dykeman, E., Benini, S., Robinson, C. V., Alonso, J. C., & Antson, A. A. (2012). Structural basis for DNA recognition and loading into a viral packaging motor. Proceedings of the National Academy of Sciences of the United States of America, 109(3), 811-816. https://doi.org/10.1073/pnas.1110270109

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title = "Structural basis for DNA recognition and loading into a viral packaging motor",

abstract = "Genome packaging into preformed viral procapsids is driven by powerful molecular motors. The small terminase protein is essential for the initial recognition of viral DNA and regulates the motor's ATPase and nuclease activities during DNA translocation. The crystal structure of a full-length small terminase protein from the Siphoviridae bacteriophage SF6, comprising the N-terminal DNA binding, the oligomerization core, and the C-terminal beta-barrel domains, reveals a nine-subunit circular assembly in which the DNA-binding domains are arranged around the oligomerization core in a highly flexible manner. Mass spectrometry analysis and four further crystal structures show that, although the full-length protein exclusively forms nine-subunit assemblies, protein constructs missing the C-terminal beta-barrel form both nine-subunit and ten-subunit assemblies, indicating the importance of the C terminus for defining the oligomeric state. The mechanism by which a ring-shaped small terminase oligomer binds viral DNA has not previously been elucidated. Here, we probed binding in vitro by using EPR and surface plasmon resonance experiments, which indicated that interaction with DNA is mediated exclusively by the DNA-binding domains and suggested a nucleosome-like model in which DNA binds around the outside of the protein oligomer.",

author = "Buettner, {Carina R.} and Maria Chechik and Miguel Ortiz-Lombardia and Callum Smits and Ima-Obong Ebong and Victor Chechik and Gunnar Jeschke and Eric Dykeman and Stefano Benini and Robinson, {Carol V.} and Alonso, {Juan C.} and Antson, {Alfred A.}",

year = "2012",

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doi = "10.1073/pnas.1110270109",

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Buettner, CR, Chechik, M, Ortiz-Lombardia, M, Smits, C, Ebong, I-O, Chechik, V, Jeschke, G, Dykeman, E, Benini, S, Robinson, CV, Alonso, JC & Antson, AA 2012, 'Structural basis for DNA recognition and loading into a viral packaging motor', Proceedings of the National Academy of Sciences of the United States of America, vol. 109, no. 3, pp. 811-816. https://doi.org/10.1073/pnas.1110270109

TY - JOUR

T1 - Structural basis for DNA recognition and loading into a viral packaging motor

AU - Buettner, Carina R.

AU - Chechik, Maria

AU - Ortiz-Lombardia, Miguel

AU - Smits, Callum

AU - Ebong, Ima-Obong

AU - Chechik, Victor

AU - Jeschke, Gunnar

AU - Dykeman, Eric

AU - Benini, Stefano

AU - Robinson, Carol V.

AU - Alonso, Juan C.

AU - Antson, Alfred A.

PY - 2012/1/17

Y1 - 2012/1/17

N2 - Genome packaging into preformed viral procapsids is driven by powerful molecular motors. The small terminase protein is essential for the initial recognition of viral DNA and regulates the motor's ATPase and nuclease activities during DNA translocation. The crystal structure of a full-length small terminase protein from the Siphoviridae bacteriophage SF6, comprising the N-terminal DNA binding, the oligomerization core, and the C-terminal beta-barrel domains, reveals a nine-subunit circular assembly in which the DNA-binding domains are arranged around the oligomerization core in a highly flexible manner. Mass spectrometry analysis and four further crystal structures show that, although the full-length protein exclusively forms nine-subunit assemblies, protein constructs missing the C-terminal beta-barrel form both nine-subunit and ten-subunit assemblies, indicating the importance of the C terminus for defining the oligomeric state. The mechanism by which a ring-shaped small terminase oligomer binds viral DNA has not previously been elucidated. Here, we probed binding in vitro by using EPR and surface plasmon resonance experiments, which indicated that interaction with DNA is mediated exclusively by the DNA-binding domains and suggested a nucleosome-like model in which DNA binds around the outside of the protein oligomer.

AB - Genome packaging into preformed viral procapsids is driven by powerful molecular motors. The small terminase protein is essential for the initial recognition of viral DNA and regulates the motor's ATPase and nuclease activities during DNA translocation. The crystal structure of a full-length small terminase protein from the Siphoviridae bacteriophage SF6, comprising the N-terminal DNA binding, the oligomerization core, and the C-terminal beta-barrel domains, reveals a nine-subunit circular assembly in which the DNA-binding domains are arranged around the oligomerization core in a highly flexible manner. Mass spectrometry analysis and four further crystal structures show that, although the full-length protein exclusively forms nine-subunit assemblies, protein constructs missing the C-terminal beta-barrel form both nine-subunit and ten-subunit assemblies, indicating the importance of the C terminus for defining the oligomeric state. The mechanism by which a ring-shaped small terminase oligomer binds viral DNA has not previously been elucidated. Here, we probed binding in vitro by using EPR and surface plasmon resonance experiments, which indicated that interaction with DNA is mediated exclusively by the DNA-binding domains and suggested a nucleosome-like model in which DNA binds around the outside of the protein oligomer.

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DO - 10.1073/pnas.1110270109

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JF - Proceedings of the National Academy of Sciences of the United States of America

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Structural basis for DNA recognition and loading into a viral packaging motor

Abstract

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