Circular permutation of DNA cytosine-N4 methyltransferases: in vivo coexistence in the BcnI system and in vitro probing by hybrid formation

Nucleic Acids Research
Giedrius VilkaitisSaulius Klimasauskas

Abstract

Sequence analysis of the BcnI restriction-modification system from Bacillus centrosporus revealed four open reading frames (bcnIC, bcnIR, bcnIB and bcnIA) that are arranged as two converging collinear pairs. One pair encodes a putative small regulatory protein, C.BcnI, and the restriction endonuclease R.BcnI. The other two gene products are the DNA cytosine-N4 methyltransferases M.BcnIA and M.BcnIB, which differ by circular permutation of conserved sequence motifs. The BcnI methyltransferases are isospecific on double-stranded DNA [methylation specificity CC(C/G)GG], but M.BcnIA can also methylate the target sites in single-stranded DNA. Functional analysis shows that bcnIA is dispensable (bcnIB is capable of protecting the DNA against the in vivo activity of bcnIR); in contrast, no stable clones were obtained if bcnIB alone was deleted from the system. By analogy with the DpnII system, the second methylase M.BcnIA may play a role in the transformation proficiency of its gram-positive host. The interchangeability of homologous elements in the beta class of cytosine-N4 methylases was probed by hybrid formation between M.BcnIB and its closest homolog M.Cfr9I (CCCGGG) employing a novel semi-random strategy combined with selection ...Continue Reading

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Citations

Jan 13, 2004·Biochemical and Biophysical Research Communications·Srivani SistlaDesirazu N Rao
Dec 6, 2011·PloS One·Tomasz P Jurkowski, Albert Jeltsch
Feb 27, 2010·Critical Reviews in Biochemistry and Molecular Biology·Urulangodi Kunhiraman Madhusoodanan, Desirazu N Rao
Apr 1, 2009·Microbiology·Beata Furmanek-BlaszkMarian Sektas
Apr 21, 2007·Journal of Molecular Biology·Monika SokolowskaMatthias Bochtler
Jun 21, 2016·Chemical Reviews·Peter Weigele, Elisabeth A Raleigh
May 31, 2018·Scientific Reports·Ewa WonsTadeusz Kaczorowski

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