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

Abstract

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 PIG
Piotr Skowron,,
- Laboratory of Genetic Engineering
, Brian P. Anton
Brian P. Anton,,
-
, Edyta Czajkowska PIG
Edyta Czajkowska ,,
- Laboratory of Genetic Engineering
, Joanna Żebrowska PIG
Joanna Żebrowska,,
- Laboratory of Genetic Engineering
, Ewa Sulecka PIG
Ewa Sulecka,,
- Laboratory of Genetic Engineering
, Daria Krefft PIG
Daria Krefft ,,
- Laboratory of Genetic Engineering
, Joanna Jeżewska-Frąckowiak PIG
Joanna Jeżewska-Frąckowiak,,
- Laboratory of Genetic Engineering
, Olga Żołnierkiewicz PIG
Olga Żołnierkiewicz,,
- Laboratory of Genetic Engineering
, Małgorzata Witkowska PIG
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
Issue year2017
Vol45
No15
Pages9005-9018
Publication size in sheets0.65
Keywords in Englishplasmids, dna, genes, nucleotides, thermus, enzymes, cytokinesis
DOIDOI:10.1093/nar/gkx599
URL https://academic.oup.com/nar/article/45/15/9005/3956632/The-third-restrictionmodification-system-from
Languageen angielski
Score (nominal)40
ScoreMinisterial score = 40.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 10.162 (2) - 2016=9.338 (5)
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