Home LiteratureArticle Details
PMID: 1480472 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

BsuBI--an isospecific restriction and modification system of PstI: characterization of the BsuBI genes and enzymes.

Nucleic acids research ·Vol. 20 ·No. 24 ·1992-12-25 ·Pages 6517-23

Xu GL, Kapfer W, Walter J, Trautner TA

Abstract

The enzymes of the Bacillus subtilis BsuBI restriction/modification (R/M) system recognize the target sequence 5'CTGCAG. The genes of the BsuBI R/M system have been cloned and sequenced and their products have been characterized following overexpression and purification. The gene of the BsuBI DNA methyltransferase (M.BsuBI) consists of 1503 bp, encoding a protein of 501 amino acids with a calculated M(r) of 57.2 kD. The gene of the restriction endonuclease (R.BsuBI), comprising 948 bp, codes for a protein of 316 amino acids with a predicted M(r) of 36.2 kD. M.BsuBI modifies the adenine (A) residue of the BsuBI target site, thus representing the first A-N6-DNA methyltransferase identified in B. subtilis. Like R.PstI, R.BsuBI cleaves between the A residue and the 3' terminal G of the target site. Both enzymes of the BsuBI R/M system are, therefore, functionally identical with those of the PstI R/M system, encoded by the Gram negative species Providencia stuartii. This functional equivalence coincides with a pronounced similarity of the BsuBI/PstI DNA methyltransferases (41% amino acid identity) and restriction endonucleases (46% amino acid identity). Since the genes are also very similar (58% nucleotide identity), the BsuBI and PstI R/M systems apparently have a common evolutionary origin. In spite of the sequence conservation the gene organization is strikingly different in the two R/M systems. While the genes of the PstI R/M system are separated and transcribed divergently, the genes of the BsuBI R/M system are transcribed in the same direction, with the 3' end of the M gene overlapping the 5' end of the R gene by 17 bp.

Related Genes
BsuBI
MeSH Terms
Amino Acid Sequence Bacillus subtilis/enzymology,genetics Base Sequence Chromosomes, Bacterial Cloning, Molecular DNA, Bacterial/genetics,isolation & purification Deoxyribonucleases, Type II Site-Specific/genetics,metabolism Escherichia coli/genetics Genes, Bacterial Genomic Library Methylation Molecular Sequence Data Oligodeoxyribonucleotides Plasmids Restriction Mapping Site-Specific DNA-Methyltransferase (Adenine-Specific)/genetics,metabolism Transcription, Genetic
Chemicals
DNA, Bacterial Oligodeoxyribonucleotides DNA modification methylase PstI Site-Specific DNA-Methyltransferase (Adenine-Specific) CTGCAG-specific type II deoxyribonucleases Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Xu G L
Max-Planck-Institut für Molekulare Genetik, Berlin, Germany.
Kapfer W
Walter J
Trautner T A
References (40)
40 references, click to expand
  1. Cloning the modification methylase gene of Bacillus sphaericus R in Escherichia coli.
    Gene. 1980 Aug;10(3):219-25 PMID: 6254840
  2. Hybridization between Escherichia coli and Shigella.
    J Bacteriol. 1957 Oct;74(4):461-76 PMID: 13475269
  3. Restriction and modification in Bacillus subtilis: two DNA methyltransferases with BsuRI specificity. I. Purification and physical properties.
    J Biol Chem. 1981 Sep 10;256(17):9340-5 PMID: 6267072
  4. Restriction and modification in Bacillus subtilis: DNA methylation potential of the related bacteriophages Z, SPR, SP beta, phi 3T, and rho 11.
    J Virol. 1983 May;46(2):446-53 PMID: 6302313
  5. Low-copy-number plasmid-cloning vectors amplifiable by derepression of an inserted foreign promoter.
    Gene. 1984 Apr;28(1):45-54 PMID: 6329915
  6. The organization and complete nucleotide sequence of the PstI restriction-modification system.
    J Biol Chem. 1984 Jun 25;259(12):8015-26 PMID: 6330092
  7. Sequence specificity of cytosine methylation in the DNA of the Chinese hamster ovary (CHO-K1) cell line.
    Biochim Biophys Acta. 1984 Dec 14;783(3):227-33 PMID: 6509058
  8. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  9. Introduction of host-controlled modification and restriction systems of Bacillus subtilis IAM1247 into Bacillus subtilis 168.
    J Bacteriol. 1979 Jul;139(1):308-10 PMID: 110786
  10. Restriction of plasmid-mediated transformation in Bacillus subtilis 168.
    Mol Gen Genet. 1979 Sep;175(2):235-7 PMID: 117281
  11. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  12. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  13. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  14. Cloning and sequencing of genes encoding the TthHB8I restriction and modification enzymes: comparison with the isoschizomeric TaqI enzymes.
    Gene. 1992 Mar 1;112(1):3-12 PMID: 1339363
  15. Characterization of the archaeal, plasmid-encoded type II restriction-modification system MthTI from Methanobacterium thermoformicicum THF: homology to the bacterial NgoPII system from Neisseria gonorrhoeae.
    J Bacteriol. 1992 Sep;174(17):5719-26 PMID: 1512204
  16. Cloning and expression of the HpaI restriction-modification genes.
    Nucleic Acids Res. 1992 Feb 25;20(4):705-9 PMID: 1542567
  17. Molecular analysis of the Bacillus subtilis bacteriophage SPP1 region encompassing genes 1 to 6. The products of gene 1 and gene 2 are required for pac cleavage.
    J Mol Biol. 1992 Mar 5;224(1):87-102 PMID: 1548711
  18. Cloning and molecular characterization of the HgiCI restriction/modification system from Herpetosiphon giganteus Hpg9 reveals high similarity to BanI.
    Eur J Biochem. 1991 Dec 18;202(3):1247-56 PMID: 1662609
  19. Cloning, characterization and evolution of the BsuFI restriction endonuclease gene of Bacillus subtilis and purification of the enzyme.
    Nucleic Acids Res. 1991 Dec 11;19(23):6457-63 PMID: 1721700
  20. Restriction and modification systems.
    Annu Rev Genet. 1991;25:585-627 PMID: 1812816
  21. Organization of restriction-modification systems.
    Nucleic Acids Res. 1991 May 25;19(10):2539-66 PMID: 2041731
  22. Sequence specificity of the human mRNA N6-adenosine methylase in vitro.
    Nucleic Acids Res. 1990 Oct 11;18(19):5735-41 PMID: 2216767
  23. Cloning, nucleotide sequence, and expression of the HincII restriction-modification system.
    Nucleic Acids Res. 1990 Jul 11;18(13):3903-11 PMID: 2374714
  24. Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants.
    Nucleic Acids Res. 1989 May 11;17(9):3469-78 PMID: 2657660
  25. Sequence motifs characteristic of DNA[cytosine-N4]methyltransferases: similarity to adenine and cytosine-C5 DNA-methylases.
    Nucleic Acids Res. 1989 Dec 11;17(23):9823-32 PMID: 2690010
  26. Cytosine-specific type II DNA methyltransferases. A conserved enzyme core with variable target-recognizing domains.
    J Mol Biol. 1989 Mar 20;206(2):305-12 PMID: 2716049
  27. Predictive motifs derived from cytosine methyltransferases.
    Nucleic Acids Res. 1989 Apr 11;17(7):2421-35 PMID: 2717398
  28. The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg.
    Mol Gen Genet. 1987 Sep;209(2):335-42 PMID: 2823077
  29. Cloning and expression of Bacillus subtilis phage DNA methyltransferase genes in Escherichia coli and B. subtilis.
    Gene. 1986;41(2-3):261-70 PMID: 3011599
  30. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector.
    Gene. 1986;48(1):119-31 PMID: 3549457
  31. Overproduction of the Bacillus sphaericus R modification methylase in Escherichia coli and its purification to homogeneity.
    Gene. 1986;50(1-3):63-7 PMID: 3556326
  32. Restriction and modification in B. subtilis. Biological aspects.
    Mol Gen Genet. 1974;131(3):181-91 PMID: 4215951
  33. Host controlled modification and restriction in Bacillus subtilis.
    Mol Gen Genet. 1974;131(4):275-80 PMID: 4215952
  34. Physical properties of the DNA of bacteriophage SP50.
    Mol Gen Genet. 1967;100(1):39-55 PMID: 4967655
  35. A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors.
    Gene. 1984 Nov;31(1-3):165-71 PMID: 6098521
  36. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis.
    Mol Gen Genet. 1982;186(3):339-46 PMID: 6181373
  37. A general method for defining restriction enzyme cleavage and recognition sites.
    Methods Enzymol. 1980;65(1):391-404 PMID: 6246360
  38. DNA protection with the DNA methylase M . BbvI from Bacillus brevis var. GB against cleavage by the restriction endonucleases PstI and PvuII.
    Gene. 1980 Jul;10(2):105-12 PMID: 6248417
  39. Properties of Bacillus subtilis 168 derivatives freed of their natural prophages.
    Gene. 1980 Dec;12(1-2):155-9 PMID: 6783474
  40. Cloning and expression of the Pst I restriction-modification system in Escherichia coli.
    Proc Natl Acad Sci U S A. 1981 Mar;78(3):1503-7 PMID: 6262807
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/20.24.6517
Published
1992-12-25
Pages
6517-23
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334566
Subset
IM
Databases
GENBANK
L01541
External Links
PubMed source DOI: 10.1093/nar/20.24.6517 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2