2009
DOI: 10.1186/1471-2199-10-52
|View full text |Cite|
|
Sign up to set email alerts
|

Cloning and analysis of a bifunctional methyltransferase/restriction endonuclease TspGWI, the prototype of a Thermus sp. enzyme family

Abstract: BackgroundRestriction-modification systems are a diverse class of enzymes. They are classified into four major types: I, II, III and IV. We have previously proposed the existence of a Thermus sp. enzyme family, which belongs to type II restriction endonucleases (REases), however, it features also some characteristics of types I and III. Members include related thermophilic endonucleases: TspGWI, TaqII, TspDTI, and Tth111II.ResultsHere we describe cloning, mutagenesis and analysis of the prototype TspGWI enzyme… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
2
1
1

Citation Types

0
62
0

Year Published

2011
2011
2019
2019

Publication Types

Select...
6
1

Relationship

2
5

Authors

Journals

citations
Cited by 24 publications
(62 citation statements)
references
References 47 publications
0
62
0
Order By: Relevance
“…Considering the fact that AdoMet is slightly inhibitory to Tsp GWI REase activity (14), we speculate that a second AdoMet binding site may exist in the Thermus family of REases/MTases. One such site would cause the allosteric stimulation of REase activity, the other would participate in the methylation reaction.…”
Section: Resultsmentioning
confidence: 99%
See 2 more Smart Citations
“…Considering the fact that AdoMet is slightly inhibitory to Tsp GWI REase activity (14), we speculate that a second AdoMet binding site may exist in the Thermus family of REases/MTases. One such site would cause the allosteric stimulation of REase activity, the other would participate in the methylation reaction.…”
Section: Resultsmentioning
confidence: 99%
“…Recombinant Tsp GWI protein was purified as described previously (14). Escherichia coli DH11S { mcrA [ mrrhsdRMS (rK, mK + )- mcrBC ] ( lac-proAB ) ( recA1398 ) deoR , rpsL , srl-thi , supE / F proAB + lacI Q Z M15 } (Life Technologies, Gaithersburg, MD, USA) was used for the transformation of ligation mixtures, DNA propagation, and tspGWIRM gene expression.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…As can be seen from Table 1, the enzymes were grouped according to their domain organisation as it was presented in Conserved Domain Database [9]. As could be judged from their domain organisation, 20 new bifunctional REases are thought to represent the fusion of a REase with MTase and target recognition subunits of the type I restriction-modification systems (R-M-S structure), having similar organisation with the known type IIC bifunctional enzymes such as AloI [10], CjeI [11], MmeI [12], PpiI [13], TstI [13] and TspGWI [14]. Type I RMS enzymes are multisubunit proteins that function as a single protein complex, consisting of R, M and S subunits [3].…”
Section: Methyltransferase Fusions With a Restriction Endonucleasementioning
confidence: 99%
“…This is perhaps why it is the only found example of natural bifunctional RMS originating from type II RMS. The newly found potential type IIC proteins are good candidates to expand on the current list of 12 bifunctional enzymes: AloI [10], BcgI [17], BseMII [18], BseRI [19], BspLU11III [20], CjeI [11], Eco57I [15], HaeIV [21], MmeI [12], PpiI [13], TstI [13] and TspGWI [14]. Taking into consideration intensiveness with what new microbial genomes have been sequencing during the last decade, new bifunctional RMS could be discovered very soon.…”
Section: Methyltransferase Fusions With a Restriction Endonucleasementioning
confidence: 99%