The third restriction-modification system from Thermus aquaticus YT-1: solving the riddle of two TaqII specificities

Piotr Skowron , Brian P. Anton , Edyta Czajkowska , Joanna Żebrowska , Ewa Sulecka , Daria Krefft , Joanna Jeżewska-Frąckowiak , Olga Żołnierkiewicz , Małgorzata Witkowska , Richard D. Morgan , Geoffrey G. Wilson , Alexey Fomenkov , Richard J. Roberts , Agnieszka Żylicz-Stachula


Two restriction–modification systems have been previously discovered in Thermus aquaticus YT-1. TaqI is a 263-amino acid (aa) Type IIP restriction enzyme that recognizes and cleaves within the symmetric sequence 5′-TCGA-3′. TaqII, in contrast, is a 1105-aa Type IIC restriction-and-modification enzyme, one of a family of Thermus homologs. TaqII was originally reported to recognize two different asymmetric sequences: 5′-GACCGA-3′ and 5′-CACCCA-3′. We previously cloned the taqIIRM gene, purified the recombinant protein from Escherichia coli, and showed that TaqII recognizes the 5′-GACCGA-3′ sequence only. Here, we report the discovery, isolation, and characterization of TaqIII, the third R–M system from T. aquaticus YT-1. TaqIII is a 1101-aa Type IIC/IIL enzyme and recognizes the 5′-CACCCA-3′ sequence previously attributed to TaqII. The cleavage site is 11/9 nucleotides downstream of the A residue. The enzyme exhibits striking biochemical similarity to TaqII. The 93% identity between their aa sequences suggests that they have a common evolutionary origin. The genes are located on two separate plasmids, and are probably paralogs or pseudoparalogs. Putative positions and aa that specify DNA recognition were identified and recognition motifs for 6 uncharacterized Thermus-family enzymes were predicted.
Author Piotr Skowron (FCh/DMBt/LGE)
Piotr Skowron,,
- Laboratory of Genetic Engineering
, Brian P. Anton
Brian P. Anton,,
, Edyta Czajkowska (FCh/DMBt/LGE)
Edyta Czajkowska ,,
- Laboratory of Genetic Engineering
, Joanna Żebrowska (FCh/DMBt/LGE)
Joanna Żebrowska,,
- Laboratory of Genetic Engineering
, Ewa Sulecka (FCh/DMBt/LGE)
Ewa Sulecka,,
- Laboratory of Genetic Engineering
, Daria Krefft (FCh/DMBt/LGE)
Daria Krefft ,,
- Laboratory of Genetic Engineering
, Joanna Jeżewska-Frąckowiak (FCh/DMBt/LGE)
Joanna Jeżewska-Frąckowiak,,
- Laboratory of Genetic Engineering
, Olga Żołnierkiewicz (FCh/DMBt/LGE)
Olga Żołnierkiewicz,,
- Laboratory of Genetic Engineering
, Małgorzata Witkowska (FCh/DMBt/LGE)
Małgorzata Witkowska,,
- Laboratory of Genetic Engineering
, Richard D. Morgan
Richard D. Morgan,,
et al.`
Other language title versions
Journal seriesNucleic Acids Research, ISSN 0305-1048, (A 40 pkt)
Issue year2017
Publication size in sheets0.65
Keywords in Englishplasmids, dna, genes, nucleotides, thermus, enzymes, cytokinesis
ASJC Classification1311 Genetics
Languageen angielski
LicenseOther; published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)40
Score sourcejournalList
ScoreMinisterial score = 40.0, 08-06-2020, ArticleFromJournal
Publication indicators WoS Citations = 1.000; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 2.657; WoS Impact Factor: 2017 = 11.561 (2) - 2017=10.235 (5)
Citation count*1 (2020-07-01)
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