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  PspGI a type II restriction endonuclease from the extreme thermophile Pyrococcus sp.: Structural and functional studies to investigate an evolutionary relationship with several mesophilic restriction enzymes

Pingoud, V., Conzelmann, C., Kinzebach, S., Sudina, A., Metelev, V., Kubareva, E., et al. (2003). PspGI a type II restriction endonuclease from the extreme thermophile Pyrococcus sp.: Structural and functional studies to investigate an evolutionary relationship with several mesophilic restriction enzymes. Journal of Molecular Biology, 329(5), 913-929. doi:10.1016/S0022-2836(03)00523-0.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-8A25-D Version Permalink: https://hdl.handle.net/11858/00-001M-0000-0010-8A26-B
Genre: Journal Article
Alternative Title : J. Mol. Biol.

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 Creators:
Pingoud, Vera, Author
Conzelmann, Charlotte, Author
Kinzebach, Steffen, Author
Sudina, Anna, Author
Metelev, Valerie, Author
Kubareva, Elena, Author
Bujnicki, Janusz M., Author
Lurz, Rudi1, Author
Lüder, Gerhild1, Author
Xu, Shuang-Yong, Author
Pingoud, Alfred, Author
Affiliations:
1Max Planck Society, ou_persistent13              

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Free keywords: restriction-modification systems; DNA binding; DNA cleavage; cross-linking; electron microscopy
 Abstract: We present here the first detailed biochemical analysis of an archaeal restriction enzyme. PspGI shows sequence similarity to SsoII, EcoRII, NgoMIV and Cfr10I, which recognize related DNA sequences. We demonstrate here that PspGI, like SsoII and unlike EcoRII or NgoMIV and Cfr10I, interacts with and cleaves DNA as a homodimer and is not stimulated by simultaneous binding to two recognition sites. PspGI and SsoII differ in their basic biochemical properties, viz. stability against chemical denaturation and proteolytic digestion, DNA binding and the pH, MgCl2 and salt-dependence of their DNA cleavage activity. In contrast, the results of mutational analyses and cross-link experiments show that PspGI and SsoII have a very similar DNA binding site and catalytic center as NgoMIV and Cfr10I (whose crystal structures are known), and presumably also as EcoRII, in spite of the fact that these enzymes, which all recognize variants of the sequence -/CC-GG- (/ denotes the site of cleavage), are representatives of different subgroups of type II restriction endonucleases. A sequence comparison of all known restriction endonuclease sequences, furthermore, suggests that several enzymes recognizing other DNA sequences also share amino acid sequence similarities with PspGI, SsoII and EcoRII in the region of the presumptive active site. These results are discussed in an evolutionary context.

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Language(s): eng - English
 Dates: 2003-06-20
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 173725
ISI: 000183671600007
DOI: 10.1016/S0022-2836(03)00523-0
 Degree: -

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Title: Journal of Molecular Biology
  Alternative Title : J. Mol. Biol.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 329 (5) Sequence Number: - Start / End Page: 913 - 929 Identifier: ISSN: 0022-2836