DOI: 10.1073/pnas.0502886102. Autoinhibition of <i>Escherichia coli</i> Rep monomer helicase activity by its 2B subdomain

Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain

10.1073/pnas.0502886102

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

DNA helicases catalyze separation of double-helical DNA into its complementary single strands, a process essential for DNA replication, recombination, and repair. The Escherichia coli Rep protein, a superfamily 1 DNA helicase, functions in DNA replication restart and is required for replication of several bacteriophages. Monomers of Rep do not display helicase activity in vitro ; in fact, DNA unwinding requires Rep dimerization. Here we show that removal of the 2B subdomain of Rep to form RepΔ2B activates monomer helicase activity, albeit with limited processivity. Although both full length Rep and RepΔ2B monomers can translocate with 3′ to 5′ directionality along single-stranded DNA, the 2B subdomain inhibits the helicase activity of full length Rep. This suggests an autoregulatory mechanism for Rep helicase, which may apply to other nonhexameric helicases, whereby helicase activity is regulated by the rotational conformational state of the 2B subdomain; formation of a Rep dimer may relieve autoinhibition by altering the 2B subdomain orientation.

Bibliography

Brendza, K. M., Cheng, W., Fischer, C. J., Chesnik, M. A., Niedziela-Majka, A., & Lohman, T. M. (2005). Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain. Proceedings of the National Academy of Sciences, 102(29), 10076â10081.

Authors 6

Katherine M. Brendza (first)
Wei Cheng (additional)
Christopher J. Fischer (additional)
Marla A. Chesnik (additional)
Anita Niedziela-Majka (additional)
Timothy M. Lohman (additional)

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Dates

Type	When
Created	20 years, 1 month ago (July 11, 2005, 8:34 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 1:35 p.m.)
Indexed	1 month, 2 weeks ago (July 16, 2025, 8:55 a.m.)
Issued	20 years, 1 month ago (July 11, 2005)
Published	20 years, 1 month ago (July 11, 2005)
Published Online	20 years, 1 month ago (July 11, 2005)
Published Print	20 years, 1 month ago (July 19, 2005)

Funders 0

None

BibTeX

@article{Brendza_2005, title={Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain}, volume={102}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0502886102}, DOI={10.1073/pnas.0502886102}, number={29}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Brendza, Katherine M. and Cheng, Wei and Fischer, Christopher J. and Chesnik, Marla A. and Niedziela-Majka, Anita and Lohman, Timothy M.}, year={2005}, month=jul, pages={10076–10081} }

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  "abstract": "<jats:p>\n            DNA helicases catalyze separation of double-helical DNA into its complementary single strands, a process essential for DNA replication, recombination, and repair. The\n            <jats:italic>Escherichia coli</jats:italic>\n            Rep protein, a superfamily 1 DNA helicase, functions in DNA replication restart and is required for replication of several bacteriophages. Monomers of Rep do not display helicase activity\n            <jats:italic>in vitro</jats:italic>\n            ; in fact, DNA unwinding requires Rep dimerization. Here we show that removal of the 2B subdomain of Rep to form Rep\u03942B activates monomer helicase activity, albeit with limited processivity. Although both full length Rep and Rep\u03942B monomers can translocate with 3\u2032 to 5\u2032 directionality along single-stranded DNA, the 2B subdomain inhibits the helicase activity of full length Rep. This suggests an autoregulatory mechanism for Rep helicase, which may apply to other nonhexameric helicases, whereby helicase activity is regulated by the rotational conformational state of the 2B subdomain; formation of a Rep dimer may relieve autoinhibition by altering the 2B subdomain orientation.\n          </jats:p>",
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