Analysis of the nucleotide and derived amino acid sequences of the SsoII restriction endonuclease and methyltransferase

Karyagina, A. S.; Лунин, В. Г.; Degtyarenko, Kirill; Uvarovc, Valentin Y.; Nikolskaya, I.I.

doi:10.1016/0378-1119(93)90756-s

Cited by 31 publications

(21 citation statements)

References 31 publications

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“…The protein sequence of Ecl18kI [11] is V99% identical to those of isoschizomeric restriction enzymes, SsoII [12], SenPI [13] and StyD4I [14]. Noteworthy, StyD4I/SsoII showed signi¢cant sequence similarity to PspGI/EcoRII endonucleases speci¢c for overlapping CCWGG sequence [15].…”

Section: Introductionmentioning

confidence: 99%

Alternative arrangements of catalytic residues at the active sites of restriction enzymes

2002

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A catalytic sequence motif PDX 10À30 (E/D)XK is found in many restriction enzymes. On the basis of sequence similarities and mapping of the conserved residues to the crystal structure of NgoMIV we suggest that residues D160, K182, R186, R188 and E195 contribute to the catalytic/DNA binding site of the Ecl18kI restriction endonuclease. Mutational analysis confirms the functional significance of the conserved residues of Ecl18kI. Therefore, we conclude that the active site motif 159 VDX 21 KX 12 E of Ecl18kI differs from the canonical PDX 10À30 (E/D)XK motif characteristic for most of the restriction enzymes. Moreover, we propose that two subfamilies of endonucleases Ecl18kI/PspGI/EcoRII and Cfr10I/Bse634I/Ngo-MIV, specific, respectively, for CCNGG/CCWGG and RCCGGY/GCCGGC sites, share conserved active site architecture and DNA binding elements. ß

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Section: Introductionmentioning

confidence: 99%

Alternative arrangements of catalytic residues at the active sites of restriction enzymes

2002

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“…It was shown that Ecll8kI enzymes recognize the 5'...$ CCNGG...3' sequence in DNA; Ec/18kl DNA methyltransferase (M.Ecll8kI) methylates the C5 carbon atom of the inner dC residue and Ec/18kI restriction endonuclease (R.Ec/18kI) cleaves DNA as shown by the arrow [1]. Genes for Ecll8kl and SsoII R-M systems have essentially the same sequence [1,2]. The nucleotide sequence data for Ec/18kI R-M system reported in [1] have been deposited in the GenBank nucleotide sequence database with accession number J16897.…”

Section: Introductionmentioning

confidence: 99%

The Ecl18kI restriction‐modification system: cloning, expression, properties of the purified enzymes

et al. 1998

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Ecll8kl is a type II restriction-modification system isolated from Enterobactev cloaceae 18kI strain. Genes encoding Ec/18kI methyltransferase (M.Ecll8kl) and Ecll8kl restriction endonuclease (R.Ecll8kI) have been cloned and expressed in Escherichia coil These enzymes recognize the 5'... $ CCNGG...3' sequence in DNA; M.Ecll8kI methylates the C5 carbon atom of the inner dC residue and R.Ec/18kI cuts DNA as shown by the arrow. The restriction endonuclease and the methyltransferase were purified from E. coli B834 [pl8Apl] cells to near homogeneity. The restriction endonuclease is present in the solution as a tetramer, while the methyltransferase is a monomer. The interactions of M.Ecl|8kI and R.Ecll8kI with 1,2-dideoxyn-ribofuranose containing DNA duplexes were investigated. The target base flipping-out mechanism is applicable in the case of M.Ecll8kI. Correct cleavage of the abasic substrates by R.Ecll8kI is accompanied by non-canonical hydrolysis of the modified strand.

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“…SsolI endonuclease encoded by pQESso9 plasmid possessed an Nterminal 6 × His affinity tail, making it possible to purify the enzyme to virtual homogeneity by one-step Ni-chelate affinity column chromatography. The introduced amino acid substitutions do not change R.SsolI recognition and cleavage properties if compared with the unmodified enzyme obtained earlier [8].…”

Section: Introductionmentioning

confidence: 65%

“…This plasmid contained the R.SsolI gene [7] in the pQE9 tac expression vector (DIAGEN GmbH, Germany). The *Corresponding author.…”

Section: Enzymes and Oligonucleotidesmentioning

confidence: 99%

Cross‐linking of SsoII restriction endonuclease to cognate and non‐cognate DNAs

et al. 1996

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Analysis of the nucleotide and derived amino acid sequences of the SsoII restriction endonuclease and methyltransferase

Cited by 31 publications

References 31 publications

Alternative arrangements of catalytic residues at the active sites of restriction enzymes

Alternative arrangements of catalytic residues at the active sites of restriction enzymes

The Ecl18kI restriction‐modification system: cloning, expression, properties of the purified enzymes

Cross‐linking of SsoII restriction endonuclease to cognate and non‐cognate DNAs

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