The NIaIV restriction and modification genes of Neisseria lactamica are flanked by leucine biosynthesis genes |
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Authors: | Peter CK Lau Farnaz Forghani Diane Labbé Hélène Bergeron Roland Brousseau and H Joachim H?ltke |
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Institution: | (1) National Research Council of Canada, Biotechnology Research Institute, 6100 Royalmount Avenue, H4P 2R2 Montreal, Quebec, Canada;(2) Department of Molecular Biology, Boehringer Mannheim GmbH, Biochemical Research Centre, Nonnenwald 2, D-8122 Penzberg, Germany |
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Abstract: | The genes encoding the Neisseria lactamica restriction endonuclease IV (R.NIaIV) and its cognate DNA methyltransferase (M.NlaIV), both of which recognize the sequence GGNNCC, have been cloned in Escherichia coli and overexpressed using the T7 polymerase/promoter system. Analysis of a sequenced 3.58 kb fragment established the gene order, leuD-M.NlaIV-R.NlaIV-leuB. The predicted primary sequence of M.NlaIV (423 amino acids) shows the highest degree of identity to a pair of cytosine-specific methyltransferases, M.BanI (44.9%) and M.HgiCI (44.3%), which recognize the sequence GGYRCC (Y, pyrimidines; R, purines). In contrast, the R.NlaIV protein sequence (243 amino acids) is unique in the existing database, a situation that holds for most endonucleases. Flanking the NlaIV modification and restriction genes are homologues of the leuD and leuB genes of enteric bacteria, which code for enzymes in the leucine biosynthesis pathway. This gene context implies a possible new mode of gene regulation for the RM.NlaIV system, which would involve a mechanism similar to the recently discovered leucine/Lrp regulon in E. coli.Abbreviations
R
purines
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Y
pyrimidines
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W
adenine or thymine
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N
any base |
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Keywords: | Endonuclease Cytosine DNA methyltransferase T7 polymerase/promoter Nucleotide sequence Lrp |
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